Difference between revisions of "HaCaT Fatty acid analysis Results"

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(Created page with "Return to HaCaT keratinocytes == Estimation of cellular arachidonic acid in unstimulated cells == Image:K06_Control-1.jpg|none|thumb|500px|Estimat...")
 
 
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[[HaCaT keratinocytes|Return to HaCaT keratinocytes]]
 
[[HaCaT keratinocytes|Return to HaCaT keratinocytes]]
  
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== Estimation of cellular arachidonic acid in unstimulated cells ==
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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 /10<sup>6</sup> cells).
  
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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.   ]]
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{| class="wikitable"
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|+  style="text-align: left;" |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/10<sup>6</sup> cells) of three independent experiments.
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|-
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! rowspan="2" style="text-align: center; font-weight:bold;" | Time (h)
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! colspan="5" style="text-align: center; font-weight:bold;" | Experiment
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|-
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| style="text-align: center; font-weight:bold;" | Unstimulated
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| style="text-align: center; font-weight:bold;" | A23187
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| style="text-align: center; font-weight:bold;" | Indomethacin + A23187
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| style="text-align: center; font-weight:bold;" | UVR
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| style="text-align: center; font-weight:bold;" | ATP
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|-
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| style="text-align: center;" | 0
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| style="text-align: center;" | 2.87 ± 0.28
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| style="text-align: center;" | 2.87 ± 0.28
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| style="text-align: center;" | 2.87 ± 0.28
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| style="text-align: center;" | 2.87 ± 0.28
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| style="text-align: center;" | 2.87 ± 0.28
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|-
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| style="text-align: center;" | 0.5
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| style="text-align: center;" | 2.68 ± 0.49
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| style="text-align: center;" | 0.28 ± 0.07
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| style="text-align: center;" | 4.18 ± 0.47
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| style="text-align: center;" | 2.41 ± 0.70
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| style="text-align: center;" | 1.27 ± 0.23
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|-
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| style="text-align: center;" | 1
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| style="text-align: center;" | 2.57 ± 1.25
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| style="text-align: center;" | 0.21 ± 0.09
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| style="text-align: center;" | 3.42 ± 0.47
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| style="text-align: center;" | 2.15 ± 0.72
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| style="text-align: center;" | 1.44 ± 0.41
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|-
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| style="text-align: center;" | 3
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| style="text-align: center;" | 2.51 ± 0.82
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| style="text-align: center;" | 0.25 ± 0.20
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| style="text-align: center;" | 1.98 ± 0.34
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| style="text-align: center;" | 1.74 ± 0.77
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| style="text-align: center;" | 1.15 ± 0.20
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|-
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| style="text-align: center;" | 6
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| style="text-align: center;" | 2.91 ± 0.22
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| style="text-align: center;" | 0.28 ± 0.35
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| style="text-align: center;" | 1.13 ± 0.49
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| style="text-align: center;" | 0.79 ± 0.82
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| style="text-align: center;" | 1.47 ± 0.28
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|}
  
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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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[[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 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.]]
 

Latest revision as of 14:08, 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).

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


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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