Difference between revisions of "HaCaT Eicosanoid Analysis Results"

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[[Image:K03_summary.jpg|none|thumb|500px|Lipid mediator levels in unstimulated (red) and stimulated with A23187 (5 µM) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.  ]]
 
[[Image:K03_summary.jpg|none|thumb|500px|Lipid mediator levels in unstimulated (red) and stimulated with A23187 (5 µM) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.  ]]
 +
 +
== ATP ==
 +
{| class="wikitable"
 +
|+  style="text-align: left;" |AA-derived lipid mediators detected in the supernatant of HaCaT keratinocytes treated with ATP (2 mM), for 0.5, 1, 3 and 6h. Analysis was performed using UPLC-ESI-MS/MS. Results are expressed mean ± SD (pg/106 cells) of three independent experiments.
 +
|-
 +
! rowspan="2" style="text-align: center; font-weight:bold;" | Time (h)
 +
! colspan="6" style="text-align: center; font-weight:bold;" | Mediator
 +
|-
 +
| style="text-align: center; font-weight:bold;" | PGE2
 +
| style="text-align: center; font-weight:bold;" | 15-keto-PGE2
 +
| style="text-align: center; font-weight:bold;" | 13,14-dihydro-15-keto-PGE2
 +
| style="text-align: center; font-weight:bold;" | PGF2α
 +
| style="text-align: center; font-weight:bold;" | 12-HETE
 +
| 15-HETE
 +
|-
 +
| style="text-align: center;" | 0
 +
| style="text-align: center;" | 1.50 ± 0.00
 +
| style="text-align: center;" | 0.37 ± 0.21
 +
| style="text-align: center;" | 2.50 ± 0.00
 +
| style="text-align: center;" | 7.50 ± 0.00
 +
| style="text-align: center;" | 1.00 ± 0.00
 +
| 2.00 ± 0.00
 +
|-
 +
| style="text-align: center;" | 0.5
 +
| style="text-align: center;" | 617.77 ± 60.96
 +
| style="text-align: center;" | 1.10 ± 0.58
 +
| style="text-align: center;" | 51.51 ± 6.47
 +
| style="text-align: center;" | 96.88 ± 13.81
 +
| style="text-align: center;" | 49.56 ± 37.94
 +
| 15.82 ± 9.10
 +
|-
 +
| style="text-align: center;" | 1
 +
| style="text-align: center;" | 849.12 ± 56.72
 +
| style="text-align: center;" | 3.60 ± 0.79
 +
| style="text-align: center;" | 86.88 ± 32.86
 +
| style="text-align: center;" | 148.10 ± 20.14
 +
| style="text-align: center;" | 32.28 ± 31.72
 +
| 10.95 ± 9.46
 +
|-
 +
| style="text-align: center;" | 3
 +
| style="text-align: center;" | 833.49 ± 170.07
 +
| style="text-align: center;" | 2.05 ± 0.65
 +
| style="text-align: center;" | 106.43 ± 12.88
 +
| style="text-align: center;" | 170.02 ± 34.73
 +
| style="text-align: center;" | 34.05 ± 39.23
 +
| 5.70 ± 7.02
 +
|-
 +
| style="text-align: center;" | 6
 +
| style="text-align: center;" | 1015.73 ± 88.65
 +
| style="text-align: center;" | 2.43 ± 1.70
 +
| style="text-align: center;" | 141.16 ± 9.62
 +
| style="text-align: center;" | 210.30 ± 43.78
 +
| style="text-align: center;" | 9.76 ± 10.52
 +
| 2.22 ± 0.54
 +
|}
 +
 +
[[Image:K04_eicosanoids.jpg|none|thumb|500px|Lipid mediator levels in unstimulated (red) and stimulated with ATP (2mM) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.  ]]
 +
 +
 +
== UVR ==
 +
 +
{| class="wikitable"
 +
|+  style="text-align: left;" |AA-derived lipid mediators detected in the supernatant of HaCaT keratinocytes treated with UVR (15 mJ/cm2), for 0.5, 1, 3 and 6h. Analysis was performed using UPLC-ESI-MS/MS. Results are expressed mean ± SD (pg/106 cells) of three independent experiments.
 +
|-
 +
! rowspan="2" style="text-align: center; font-weight:bold;" | Time (h)
 +
! colspan="6" style="text-align: center; font-weight:bold;" | Mediator
 +
|-
 +
| style="text-align: center; font-weight:bold;" | PGE2
 +
| style="text-align: center; font-weight:bold;" | 15-keto-PGE2
 +
| style="text-align: center; font-weight:bold;" | 13,14-dihydro-15-keto-PGE2
 +
| style="text-align: center; font-weight:bold;" | PGF2α
 +
| style="text-align: center; font-weight:bold;" | 12-HETE
 +
| style="font-weight:bold;" | 15-HETE
 +
|-
 +
| style="text-align: center;" | 0
 +
| style="text-align: center;" | 203.29 ± 315.50
 +
| style="text-align: center;" | 0.50 ± 0.00
 +
| style="text-align: center;" | 2.00 ± 0.00
 +
| style="text-align: center;" | 0.00 ± 0.00
 +
| style="text-align: center;" | 3.50 ± 0.00
 +
| 2.30 ± 0.00
 +
|-
 +
| style="text-align: center;" | 0.5
 +
| style="text-align: center;" | 1.50 ± 0.00
 +
| style="text-align: center;" | 0.75 ± 0.39
 +
| style="text-align: center;" | 2.00 ± 0.00
 +
| style="text-align: center;" | 88.72 ± 33.06
 +
| style="text-align: center;" | 135.07 ± 142.32
 +
| 40.09 ± 35.24
 +
|-
 +
| style="text-align: center;" | 1
 +
| style="text-align: center;" | 69.26 ± 60.99
 +
| style="text-align: center;" | 1.05 ± 0.99
 +
| style="text-align: center;" | 2.29 ± 1.56
 +
| style="text-align: center;" | 90.56 ± 11.96
 +
| style="text-align: center;" | 131.60 ± 100.39
 +
| 46.54 ± 35.33
 +
|-
 +
| style="text-align: center;" | 3
 +
| style="text-align: center;" | 1.50 ± 0.00
 +
| style="text-align: center;" | 9.61 ± 7.15
 +
| style="text-align: center;" | 3.53 ± 2.41
 +
| style="text-align: center;" | 222.44 ± 6.73
 +
| style="text-align: center;" | 103.71 ± 78.12
 +
| 58.45 ± 43.57
 +
|-
 +
| style="text-align: center;" | 6
 +
| style="text-align: center;" | 5667.91 ± 4967.29
 +
| style="text-align: center;" | 152.58 ± 51.19
 +
| style="text-align: center;" | 6.21 ± 4.30
 +
| style="text-align: center;" | 1657.38 ± 297.77
 +
| style="text-align: center;" | 766.69 ± 206.41
 +
| 648.45 ± 219.84
 +
|}
 +
 +
[[Image:K01_eicosanoids.jpg|none|thumb|500px|Lipid mediator levels in unstimulated (red) and irradiated (15 mJ/cm2) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE.  The data presented is from three independent experiments, each with a technical replicate.]]
 +
 +
== Calcium ionophore + Indomethacin ==
 +
 +
{| class="wikitable"
 +
|+  style="text-align: left;" | AA-derived lipid mediators detected in the supernatant of COX inhibited HaCaT keratinocytes treated with A23187 (5 µM), for 0.5, 1, 3 and 6h. Analysis was performed using UPLC-ESI-MS/MS. Results are expressed mean ± SD (pg/106 cells) of three independent experiments.
 +
|-
 +
! rowspan="2" style="text-align: center; font-weight:bold;" | Time (h)
 +
! colspan="6" style="text-align: center; font-weight:bold;" | Mediator
 +
|-
 +
| style="text-align: center; font-weight:bold;" | PGE2
 +
| style="text-align: center; font-weight:bold;" | 15-keto-PGE2
 +
| style="text-align: center; font-weight:bold;" | 13,14-dihydro-15-keto-PGE2
 +
| style="text-align: center; font-weight:bold;" | PGF2α
 +
| style="text-align: center; font-weight:bold;" | 12-HETE
 +
| 15-HETE
 +
|-
 +
| style="text-align: center;" | 0
 +
| style="text-align: center;" | 26.81 ± 10.56
 +
| style="text-align: center;" | 0.50 ± 0.00
 +
| style="text-align: center;" | 2.00 ± 0.00
 +
| style="text-align: center;" | 7.00 ± 0.00
 +
| style="text-align: center;" | 0.29 ± 0.00
 +
| 1.92 ± 0.94
 +
|-
 +
| style="text-align: center;" | 0.5
 +
| style="text-align: center;" | 34.23 ± 22.16
 +
| style="text-align: center;" | 0.50 ± 0.00
 +
| style="text-align: center;" | 1.88 ± 0.57
 +
| style="text-align: center;" | 7.00 ± 0.00
 +
| style="text-align: center;" | 7.20 ± 10.94
 +
| 29.71 ± 60.25
 +
|-
 +
| style="text-align: center;" | 1
 +
| style="text-align: center;" | 25.85 ± 9.25
 +
| style="text-align: center;" | 0.50 ± 0.00
 +
| style="text-align: center;" | 32.91 ± 28.56
 +
| style="text-align: center;" | 7.00 ± 0.00
 +
| style="text-align: center;" | 141.09 ± 24.66
 +
| 54.23 ± 51.50
 +
|-
 +
| style="text-align: center;" | 3
 +
| style="text-align: center;" | 4731.85 ± 7300.78
 +
| style="text-align: center;" | 2266.13 ± 371.89
 +
| style="text-align: center;" | 1817.62 ± 907.18
 +
| style="text-align: center;" | 3268.46 ± 658.45
 +
| style="text-align: center;" | 17918.03 ± 8595.61
 +
| 55405.98 ± 17910.03
 +
|-
 +
| style="text-align: center;" | 6
 +
| style="text-align: center;" | 4843.70 ± 7474.03
 +
| style="text-align: center;" | 1620.01 ± 317.98
 +
| style="text-align: center;" | 900.50 ± 199.18
 +
| style="text-align: center;" | 4754.66 ± 596.47
 +
| style="text-align: center;" | 61471.62 ± 16768.21
 +
| 105543.44 ± 41314.43
 +
|}
 +
 +
 +
[[Image:K07_eicosanoids.jpg|none|thumb|500px|Lipid mediator levels in unstimulated (red) and stimulated with A23187 (5 µM) following 1h incubation with indomethacin (10 mM) (blue) HaCaT keratinocytes, over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE.  The data presented is from three independent experiments, each with a technical replicate. ]]

Revision as of 15:32, 31 May 2019

Return to overview


Calcium Ionophore

AA-derived lipid mediators detected in the supernatant of HaCaT keratinocytes treated with A23187 (5 µM), for 0.5, 1, 3 and 6h. Analysis was performed using UPLC-ESI-MS/MS. Results are expressed mean ± SD (pg/106 cells) of three independent experiments.
Time (h) Mediator
PGE2 15-keto-PGE2 13,14-dihydro-15-keto-PGE2 PGF2α 12-HETE 15-HETE
0 9.45 ± 7.56 0.50 ± 0.00 2.00 ± 0.00 7.00 ± 0.00 6.44 ± 4.40 3.72 ± 1.08
0.5 601.26 ± 162.02 65.52 ± 57.95 211.38 ± 61.05 106.06 ± 39.89 19.49 ± 10.36 163.26 ± 143.10
1 14371.34 ± 858.76 2136.87 ± 588.92 2461.84 ± 452.79 2787.86 ± 489.04 363.25 ± 96.91 4050.14 ± 2998.16
3 79576.88 ± 15355.19 12230.83 ± 2179.66 11025.93 ± 2287.76 19276.46 ± 3732.43 4872.78 ± 849.95 41799.69 ± 31552.30
6 111115.14 ± 32976.86 18619.32 ± 5691.99 15799.31 ± 5867.25 34965.99 ± 11250.10 7156.92 ± 1550.86 55738.98 ± 40130.05


Lipid mediator levels in unstimulated (red) and stimulated with A23187 (5 µM) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.

ATP

AA-derived lipid mediators detected in the supernatant of HaCaT keratinocytes treated with ATP (2 mM), for 0.5, 1, 3 and 6h. Analysis was performed using UPLC-ESI-MS/MS. Results are expressed mean ± SD (pg/106 cells) of three independent experiments.
Time (h) Mediator
PGE2 15-keto-PGE2 13,14-dihydro-15-keto-PGE2 PGF2α 12-HETE 15-HETE
0 1.50 ± 0.00 0.37 ± 0.21 2.50 ± 0.00 7.50 ± 0.00 1.00 ± 0.00 2.00 ± 0.00
0.5 617.77 ± 60.96 1.10 ± 0.58 51.51 ± 6.47 96.88 ± 13.81 49.56 ± 37.94 15.82 ± 9.10
1 849.12 ± 56.72 3.60 ± 0.79 86.88 ± 32.86 148.10 ± 20.14 32.28 ± 31.72 10.95 ± 9.46
3 833.49 ± 170.07 2.05 ± 0.65 106.43 ± 12.88 170.02 ± 34.73 34.05 ± 39.23 5.70 ± 7.02
6 1015.73 ± 88.65 2.43 ± 1.70 141.16 ± 9.62 210.30 ± 43.78 9.76 ± 10.52 2.22 ± 0.54
Lipid mediator levels in unstimulated (red) and stimulated with ATP (2mM) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.


UVR

AA-derived lipid mediators detected in the supernatant of HaCaT keratinocytes treated with UVR (15 mJ/cm2), for 0.5, 1, 3 and 6h. Analysis was performed using UPLC-ESI-MS/MS. Results are expressed mean ± SD (pg/106 cells) of three independent experiments.
Time (h) Mediator
PGE2 15-keto-PGE2 13,14-dihydro-15-keto-PGE2 PGF2α 12-HETE 15-HETE
0 203.29 ± 315.50 0.50 ± 0.00 2.00 ± 0.00 0.00 ± 0.00 3.50 ± 0.00 2.30 ± 0.00
0.5 1.50 ± 0.00 0.75 ± 0.39 2.00 ± 0.00 88.72 ± 33.06 135.07 ± 142.32 40.09 ± 35.24
1 69.26 ± 60.99 1.05 ± 0.99 2.29 ± 1.56 90.56 ± 11.96 131.60 ± 100.39 46.54 ± 35.33
3 1.50 ± 0.00 9.61 ± 7.15 3.53 ± 2.41 222.44 ± 6.73 103.71 ± 78.12 58.45 ± 43.57
6 5667.91 ± 4967.29 152.58 ± 51.19 6.21 ± 4.30 1657.38 ± 297.77 766.69 ± 206.41 648.45 ± 219.84
Lipid mediator levels in unstimulated (red) and irradiated (15 mJ/cm2) (blue) HaCaT keratinocytes over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.

Calcium ionophore + Indomethacin

AA-derived lipid mediators detected in the supernatant of COX inhibited HaCaT keratinocytes treated with A23187 (5 µM), for 0.5, 1, 3 and 6h. Analysis was performed using UPLC-ESI-MS/MS. Results are expressed mean ± SD (pg/106 cells) of three independent experiments.
Time (h) Mediator
PGE2 15-keto-PGE2 13,14-dihydro-15-keto-PGE2 PGF2α 12-HETE 15-HETE
0 26.81 ± 10.56 0.50 ± 0.00 2.00 ± 0.00 7.00 ± 0.00 0.29 ± 0.00 1.92 ± 0.94
0.5 34.23 ± 22.16 0.50 ± 0.00 1.88 ± 0.57 7.00 ± 0.00 7.20 ± 10.94 29.71 ± 60.25
1 25.85 ± 9.25 0.50 ± 0.00 32.91 ± 28.56 7.00 ± 0.00 141.09 ± 24.66 54.23 ± 51.50
3 4731.85 ± 7300.78 2266.13 ± 371.89 1817.62 ± 907.18 3268.46 ± 658.45 17918.03 ± 8595.61 55405.98 ± 17910.03
6 4843.70 ± 7474.03 1620.01 ± 317.98 900.50 ± 199.18 4754.66 ± 596.47 61471.62 ± 16768.21 105543.44 ± 41314.43


Lipid mediator levels in unstimulated (red) and stimulated with A23187 (5 µM) following 1h incubation with indomethacin (10 mM) (blue) HaCaT keratinocytes, over 6h. Where (A) PGE2, (B) 15-keto-PGE2, (C) 13,14-dihydro-15-keto-PGE2, (D) PGF2α, (E) 12-HETE and (D) 15-HETE. The data presented is from three independent experiments, each with a technical replicate.
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