HaCaT Fatty acid analysis Results

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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 /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).