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. 2022 Apr 23;23(9):4666. doi: 10.3390/ijms23094666

Figure 5.

Figure 5

Figure 5

Involvement of ER stress-regulated signaling in norketamine (NK)-induced RT4 cell apoptosis. Cells were treated with NK (0.2 mM) for different time intervals (2–24 h), and the levels of protein expression for (A) GRP78, GRP94, CHOP, XBP-1s, ATF-4, cleaved ATF-6, and caspase-12, and (B) the phosphorylated or total eIF2α, PERK, and IRE-1 were examined using Western blot analysis. Additionally, RT4 cells were treated with NK (0.2 mM) for 24 h in the absence or presence of an ER stress inhibitor 4-phenylbutyric acid (4-PBA; 2 mM; for 1 h prior to treated with NK), and (C) the levels of protein expression for GRP78, CHOP, XBP-1s, ATF-4, cleaved ATF-6, Caspase-3, -7, -12, and PARP were examined using Western blot analysis; and (D) caspase-3 activity was detected using the caspase-3 Activity Assay Kit. Results shown in A, B, and C on representative images, and quantification was determined by densitometric analysis. Each bar presented is the mean ± SD of three independent experiments. Data in D are presented as the means ± SD of four independent experiments assayed in triplicate. * p < 0.05 compared to vehicle control. # p < 0.05 compared to NK treatment alone.