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. 2021 Sep 18;10(9):1492. doi: 10.3390/antiox10091492

Figure 4.

Figure 4

Fisetin inhibits PM2.5-induced apoptosis by alleviating ER stress. HaCaT keratinocytes were treated with fisetin (0–10 µM) for 2 h and subsequently exposed to 100 µg/mL PM2.5 for 24 h. (A) The total proteins were extracted, and Western blotting was performed for detecting the expression of GRP78, p-eIF2α, eIF2α, ATF4, and CHOP. β-Actin was used as the loading control. (B) The cells were treated with 10 µM fisetin or 20 µM salubrinal in the presence or absence of 100 µg/mL PM2.5 for 24 h. The cells were incubated with Ca2+-sensitive Fluo-4 AM for 10 min and live images were captured using a CELENA S Digital Imaging System. Scale bar = 100 µm. (C,D) In a parallel experiment, the cells were treated with 10 µM fisetin or 20 µM salubrinal in the presence or absence of 100 µg/mL PM2.5 for 24 h. The cells were stained with a (C) Muse Oxidative Stress Assay Kit and (D) Muse Annexin V & Dead Cell Assay Kit. *** p < 0.001 vs. untreated cells and # p < 0.05 vs. PM2.5-treated cells.