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− | == Estimation of cellular arachidonic acid in unstimulated cells == | + | == Summary == |
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− | [[Image:K06_Control-1.jpg|none|thumb|500px|Estimation of AA in unstimulated HaCaT keratinocytes over 6h. The data presented is from three independent experiments. ]] | + | [[Image:Hacat_AA.jpg|none|thumb|500px|Levels of cellular arachidonic acid in HaCaT keratinocytes following stimulation with (A) A23187, (B) UVR and (C) ATP 6h post stimulation. Data points are from n=3 independent experiments. Non-linear lines of best fit are shown for untreated control cells (red), treated cells (blue) and treated with indomethacin and A23187 cells (black). Abbreviations: control cells (CTR), calcium ionophore (A23187, 5 µM) (CaI) and indomethacin (10 mM, 1h) (IND). ]] |
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− | == Estimation of cellular arachidonic acid following stimulation with calcium ionophore (A23187) ==
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− | [[Image:K03-1.jpg|none|thumb|500px|Estimation of AA in HaCaT keratinocytes stimulated with A23187 (5 µM) for 6h. The data presented is from three independent experiments. ]]
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− | == Estimation of cellular arachidonic acid following stimulation with ATP ==
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− | [[Image:K04.jpg|none|thumb|500px|Estimation of AA in HaCaT keratinocytes stimulated with ATP (2 mM) for 6h. The data presented is from three independent experiments.]]
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− | == Estimation of cellular arachidonic acid following stimulation with UVR ==
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− | [[Image:K01-1.jpg|none|thumb|500px|Estimation of AA in HaCaT keratinocytes over 6h following stimulation with UVR (15 mJ/cm2). The data presented is from three independent experiments.]]
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− | == Estimation of cellular arachidonic acid following inhibition with indomethacin ==
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− | [[Image:K07-1.jpg|none|thumb|500px|Estimation of AA in HaCaT keratinocytes inhibited with indomethacin (10 mM, 1h) and stimulated with A23187 (5 µM) for 6h. The data presented is from three independent experiments.]]
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Revision as of 14:07, 26 August 2019
Return to HaCaT keratinocytes
The aim of this study was to estimate the levels of AA in cells which were unstimulated and stimulated with A23187 (with/without COX inhibition), ATP and UVR. GC-FID was used for this fatty acid analysis and the values were normalized using cell numbers (ug /106 cells).
Summary Table
The estimated AA levels in HaCaT keratinocytes (unstimulated, A23187 (5µM), COX inhibited and A23187 (5 µM), ATP (2 mM) and UVR (15 mJ/cm2)) for 0.5, 1, 3 and 6h. Analysis was performed using GC-FID. Results are expressed mean ± SD (ug/106 cells) of three independent experiments.
Time (h)
|
Experiment
|
Unstimulated
|
A23187
|
Indomethacin + A23187
|
UVR
|
ATP
|
0
|
2.87 ± 0.28
|
2.87 ± 0.28
|
2.87 ± 0.28
|
2.87 ± 0.28
|
2.87 ± 0.28
|
0.5
|
2.68 ± 0.49
|
0.28 ± 0.07
|
4.18 ± 0.47
|
2.41 ± 0.70
|
1.27 ± 0.23
|
1
|
2.57 ± 1.25
|
0.21 ± 0.09
|
3.42 ± 0.47
|
2.15 ± 0.72
|
1.44 ± 0.41
|
3
|
2.51 ± 0.82
|
0.25 ± 0.20
|
1.98 ± 0.34
|
1.74 ± 0.77
|
1.15 ± 0.20
|
6
|
2.91 ± 0.22
|
0.28 ± 0.35
|
1.13 ± 0.49
|
0.79 ± 0.82
|
1.47 ± 0.28
|
Summary
Levels of cellular arachidonic acid in HaCaT keratinocytes following stimulation with (A) A23187, (B) UVR and (C) ATP 6h post stimulation. Data points are from n=3 independent experiments. Non-linear lines of best fit are shown for untreated control cells (red), treated cells (blue) and treated with indomethacin and A23187 cells (black). Abbreviations: control cells (CTR), calcium ionophore (A23187, 5 µM) (CaI) and indomethacin (10 mM, 1h) (IND).