Fig. 3: TRPML1 is a gravity-sensitive receptor.

A The RT-qPCR analysis for the mRNA level of TRPML1 in HepaRG cells without or with RFC (10 rpm/min) for 72 h. Mean ± SD (n = 3) by Student’s t-test. B Western blotting analysis for the protein content of TRPML1 in HepaRG cells incubated without or with RFC. C The RT-qPCR and western blotting assays were conducted to analyze the transfection efficiency of shTRPML1. Mean ± SD (n = 3) by Student’s t-test. D Immunofluorescence staining of AFP, ALB, CK18 (red), and DAPI (blue) in HepaRG cells transfected without or with shTRPML1 (2.5 µM) for 72 h, under RFC conditions or not (Con: the undifferentiated group, the other groups: differentiated with DMSO). Scale bar: 80 μm. Mean ± SD (n = 5) by ANOVA. E The RT-qPCR and western blotting assays were conducted to analyze the transfection efficiency of siTRPML1. Mean ± SD (n = 3) by Student’s t-test. F Immunofluorescence staining of AFP, ALB, CK18 (red) and DAPI (blue) in HepaRG cells transfected without or with siTRPML1 (2.5 µM) for 72 h under RFC conditions. Scale bar: 80 μm. Mean ± SD by Student’s t-test. G The RT-qPCR analysis of AFP, ALB, CK18 in HepaRG cells transfected without or with siTRPML1 (2.5 µM) for 72 h under RFC conditions. Mean ± SD by Student’s t-test. H The Western blot analysis of AFP, ALB, and CK18 in HepaRG cells transfected without or with siTRPML1 (2.5 µM) for 72 h under RFC conditions. Mean ± SD by Student’s t-test. The data are based on three independent experiments. *P < 0.05, **P < 0.01.