Additional supporting data related to Fig. 1 . (A) SOCE in thapsigargin (TG)-treated Jurkat cells was measured by flow cytometry. (B) Jurkat cells were treated with DMSO, TG (10 nM), or TG (10 nM) plus CsA (1 μM) for 48 h. The expression of PD-1 was examined by flow cytometry. Mean fluorescence intensities of PD-1 staining are shown; FMO is unstained control. (C) Jurkat cells were treated with DMSO or indicated concentrations of TG for 48 h. PD-1 expression was examined. Mean fluorescence intensities of PD-1 staining are shown; FMO is unstained control. (D) Jurkat cells were treated as in C. The percentage of live cells was determined by flow cytometry with DAPI staining. (E) The expression of VMP1 in Jurkat cells expressing indicated sgRNAs and Cas9 was analyzed by immunoblot. (F) TG-induced Ca²⁺ influx in Jurkat cells expressing indicated sgRNA and Cas9 was examined by flow cytometry, and the expression of ATG5 was analyzed by immunoblot. (G) The expression of VMP1 in HEK293T cells expressing indicated sgRNAs, and Cas9 was analyzed by immunoblot. (H) Validation of monoclonal VMP1-KO HEK293T cells by immunoblot. (I) ER Ca²⁺ and SOCE in VMP1-KO cells were examined by flow cytometry. (J) The expression of VMP1 in HEK293T-G-CEPIA1er cells expressing indicated sgRNAs and Cas9 was analyzed by immunoblot. (K) 1 × 10⁵ HEK293T cells expressing indicated sgRNAs and Cas9 were plated in 12-well plates. 24 h later, cells were treated with DMSO or TG (10 nM) for another 48 h. (A–D and F) Representative plots from one of three independent experiments. (E–H and J) Representative blots from one of two independent experiments. (I and K) Representative plots and statistics from one of three independent experiments. n = 3 in I and K. Each symbol represents one sample; data represent mean ± SEM, one-way ANOVA in I, two-tailed unpaired t test in K; * <0.05, ** <0.01, **** <0.001. Source data are available for this figure: SourceData FS1.