Fig. 2: AlphaSec31AKD cells show survival phenotype against ER stress.

A Generation of alphaSec31A KD cells. Left: Immunoblots of control cells and alphaSec31A cells. Quantification is shown in the right panel. Data are presented as mean values ± SEM. (two-tailed Student’s t test). **p < 0.01. Three biological replicates cultured in independent wells were examined. B Immunoblots of control and αSec31A knockdown cells treated with thapsigargin (TG, 1 μM) for 5 h. Quantified band intensities are presented as mean ± SEM. One-way ANOVA revealed a significant overall difference among groups (p = 0.0001), and Tukey’s post-hoc test was used to assess pairwise significance.*p < 0.05, **p < 0.01. Four biological replicates cultured in independent wells were examined. C Cell viability assays of control and alphaSec31A cells. n = 6 (0 h, 48 h), n = 4 (63 h), each representing an independent biological replicate. Data are presented as mean values ± SEM. (two-tailed Student’s t test). *p < 0.05. D Immunoblots of cleaved caspase-3 after treating control and alphaSec31A cells with thapsigargin (TG), tunicamycin (Tm), and Brefeldin A (BA) for 5 h. Figure E, F Immunoblots of control and αSec31A cells treated with TNFα (1000 U/ml) and IFN-γ (1000 U/ml) (E) or palmitate (500 μM) for 16 h (F). G Immunoblots of control and siRNA-mediated Sec31A knockdown cells treated with TG (1 μM) for 5 h. H Cell viability of control and αSec31A KD cells after concentration-dependent H₂O₂ treatment (n = 3, each an independent biological replicate). Data are presented as mean values ± SEM. (two-tailed Student’s t test). *p < 0.05. I, J Flow cytometric analysis of apoptosis in control and αSec31A knockdown cells treated with thapsigargin (TG, 1 μM for 5 h) and following 24 h of TG removal (n = 3 independent biological replicates). I Representative quadrant dot plots (J) Quantification of cell populations across quadrants (Q1–Q4) under control, TG, and post-TG recovery conditions. Data are presented as mean ± SEM. Two-way ANOVA revealed significant main and/or interaction effects, and Tukey’s post-hoc test was applied to assess pairwise differences. *p  <  0.05, **p  <  0.01,***p  <  0.001. Data are presented as mean values ± SEM. Source data are provided as a Source Data file.