Fig 2: Activation of PSCs impaired upon CM5480 treatment in vitro.(A) Representative scheme of in vitro culturing of mouse PSC culture marking the CM5480 treatments during the 1-week time course. (B) Immunofluorescence staining and relative intensity values of VIM, Orai1, GFAP, and a-SMA in cultured PSCs. Scale bar: 100 µm. (C) Representative images and ratio of Oil Red O+ cells in cultured PSCs. n = 4/group, 5–7 images/culture, data points in the bars indicate individual field of views (B and C). Scale bar: 20 µm. (D) mRNA expression of Orai1, Acta2, Tgfb1, Saraf, and Fn1 in cultured PSCs (n = 4/group from 4 animals). (E) Representative images and relative intensity values of Orai1 and percentage of PSCs with spontaneous Orai1 puncta formation in the control culture (n = 4/group, 5–7 images/culture). Scale bar: 10 µm. A P value less than 0.05 was considered significant by unpaired t test (B and D [Orai1, Fn1], and E) and Mann-Whitney U test (C and D [Saraf, Tgfb1, Acta2]). Data represent mean ± SEM. Explanatory images were created with BioRender.

Fig 3: Orai1 inhibition impairs the activation of PSCs in CP.(A) Immunofluorescence staining and relative intensity values of a-SMA and VIM in mouse pancreatic tissue (n = 3/group, 5–7 images/animal). Scale bar: 20 µm. (B) Acta2 gene expression in pancreatic tissue (n = 3/group). (C) IHC images of a-SMA (top) and GFAP (bottom) in mouse pancreatic tissues (n = 3/group, 5–7 images/animal). Scale bar: 50 µm. Bar charts summarize the ratio of a-SMA+ and GFAP+ and GFAP/a-SMA–double-positive cells (activated PSCs) (orange bars) in percentage of GFAP+ total cell number (bars with dashed line). A P value less than 0.05 was considered significant by unpaired t test (A) and Mann-Whitney U test (B and C). Data represent mean ± SEM. Explanatory images were created with BioRender. cer., cerulein; p.s., physiological saline.