Fig. 2

Inhibition of TRPM7, ORAI1 and STIM1 reduces cell migration and invasion in MM cells likely in concomitant with a decreased cellular O-GlcNAcylation. a–c Western blot analysis showing TRPM7, ORAI1, and STIM1 knockdown efficiency in sgTRPM7, sgORAI1 and sgSTIM1 RPMI8226 cells, respectively. d–f Cell migration and invasion were evaluated by Transwell assays at 48 h, where the penetrating cells were stained by Hoechst 33342 dye. (upper) Representative micrographs of migrating/invading cells are shown. Scale bar = 100 μm. (lower) Quantitative data showing the percentage of migrating/invading cells relative to WT control cells. Data mean ± SD (n = 4). ***P < 0.001 versus WT; two-tailed Student’s t-test. g RPMI8226 cells were treated with 2-APB (0–50 μM), AnCoA4 (0–40 μM), and SKF96365 (0–40 μM) for 24 h, and cellular O-GlcNAcylation level was evaluated by Western blotting. h Quantitative analysis of O-GlcNAcylation level in treated cells, normalized to β-actin and relative to non-treated cells (NTX). Data are mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001 versus NTX; two-tailed Student’s t-test. i–k Cellular O-GlcNAcylation level was compared between sgTRPM7, sgORAI1 or sgSTIM1 and WT RPMI8226 cells. Data are mean ± SD (n = 4). *P < 0.05, ***P < 0.001 versus NTX; two-tailed Student’s t-test