Fig. 2

TMCO1 KD enhances IRE1α-XBP1 pathway in a Ca.²⁺ dependent manner. a Western blotting analysis of p-IRE1α, IRE1α and XBP1s levels in WT and TMCO1 KD cells treated with/without 50 μM BAPTA-AM for 2 h. Relative quantification of p-IRE1α level is shown in the right panel. Data are shown as the mean protein intensity normalized to GAPDH ± SEM from 3 independent experiments. **P < 0.01. b qRT-PCR analysis of Xbp1 mRNA splicing in WT and TMCO1 KD cells treated with 3 μg/ml TM along with/without 50 μM BAPTA-AM for 4 h. Bar graphs represent the mean ± SEM from three independent assays. *P < 0.05; **P < 0.01. c The levels of XBP1s in WT and TMCO1 KD cells were measured by western blotting analysis after the treatment of 3 μg/ml TM along with/without 50 μM BAPTA-AM for 4 h. GAPDH was used as a loading control. Relative quantification of XBP1s levels was shown in the right panel. Data are shown as the mean protein intensity normalized to GAPDH ± SEM from 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. d qRT-PCR analysis of Xbp1 mRNA splicing in WT and TMCO1 KD cells treated with 1 μM TG along with/without 50 μM BAPTA-AM for 4 h. Bar graphs represent the mean ± SEM from three independent assays. **P < 0.01, ***P < 0.001. e The levels of XBP1s in WT and TMCO1 KD cells were measured (Left) and quantified (Right) by western blotting after the treatment of 1 μM TG along with/without 50 μM BAPTA-AM for 4 h. GAPDH was used as a loading control. Data are shown as the mean protein intensity normalized to GAPDH ± SEM from 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. f qRT-PCR analysis of Xbp1 mRNA splicing in WT and TMCO1 KD cells treated with 5 mM DTT along with/without 50 μM BAPTA-AM for 4 h. Bar graphs represent the mean ± SEM from three independent assays. *P < 0.05; **P < 0.01, ***P < 0.001. g The levels of XBP1s in WT and TMCO1 KD HeLa cells were measured (Left) and quantified (Right) by western blotting analysis after the treatment of 5 mM DTT along with/without 50 μM BAPTA-AM for 4 h. GAPDH was used as a loading control. Data are shown as the mean protein intensity normalized to GAPDH ± SEM from 3 independent experiments. **P < 0.01, ***P < 0.001