Fig 7. TJ-35 suppresses cytosolic Ca²⁺ increment under starvation condition.

A. HeLa cells were cultured with DMEM for 24 h, shifted to EBSS for 2 h. The cells were stained with Fluo-8 for 30 min. HeLa cells were transiently transfected with G-CaMP3 (Addgene: 22692) for 24 h, and shifted to EBSS for 2 h. For measuring Fmax, ionomycin was added to DMEM. For measuring Fmin, HeLa cells transiently transfected with G-CaMP3 were cultured and added ionomycin with BAPTA in HBSS. B. HeLa cells were cultured with DMEM for 24 h, shifted to EBSS with or without TJ-35. The cells were stained with Fluo-8 for 30 min. We counted 30 cells in three independent experiments. C. HeLa cells were transiently transfected with G-CaMP3 for 24 h, shifted to EBSS with or without TJ-35. We counted 30 cells in three independent experiments. ABC. Fluorescence intensity in ROI within cytoplasm was measured. Median: line; upper and lower quartiles: boxes; 1.5-interquartile range: whiskers. * denotes p<0.05 (unpaired two-tailed Student’s t-test) between EBSS and EBSS plus TJ-35.