Difference between revisions of "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 /10<sup>6</sup> cells).
 
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).
 
== Summary Table ==
 
  
 
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== Estimation of cellular arachidonic acid in unstimulated cells ==
 
 
[[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.  ]]
 
 
== Estimation of cellular arachidonic acid following stimulation with calcium ionophore (A23187) ==
 
 
[[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.  ]]
 
 
== Estimation of cellular arachidonic acid following stimulation with ATP ==
 
 
[[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.]]
 
 
 
== Estimation of cellular arachidonic acid following stimulation with UVR ==
 
 
[[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.]]
 
 
== Estimation of cellular arachidonic acid following inhibition with indomethacin ==
 
  
[[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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[[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).  ]]

Latest revision as of 13:08, 26 August 2019

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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).