Figure 4
From: Molecular pathway of near-infrared laser phototoxicity involves ATF-4 orchestrated ER stress
Laser induces ER stress at sub-phototoxic doses.
(a) ER stress markers after varying doses of laser treatment were assessed at 24 hrs using immunoblotting. (b) Localization of ATF 4 in HaCaT cells were assessed after 6 hrs following laser treatment. Scale bars = 200 μm. (c) Level of autophagy (LC3A/B-ii/i), ATF-4 and HSP70 were assessed by immunoblotting after 24 hrs of laser treatment. HaCaT cells were pretreated with HSP inhibitors (d) autophagy inhibitors (e) or pre-treated with LPS (2 ng/ml), PMA (1X) and TMFC (Tamoxifen Citrate 40 μM) (f) for 2 hrs and were then challenged with laser treatment and cell viability was assessed at 24 hrs. Significance based on one-way ANOVA with the respective controls (n = 3). HaCaT and NOKSI cells were pretreated with varying concentrations of Rapamycin and challenged with phototoxic dose followed by the assessment of cellular viability (g) and immunoblotting (h) Images in a gel are cropped (horizontally) and placed together for better clarity of results and were run in the same experimental conditions. Significance was determined using two-way ANOVA among different treatments using the Bonferroni’s multiple comparison test. Statistical significance are indicated as P < 0.0005 (***), <0.00001(****) and not significant (n.s.).
