Back to article: The primary cilium protein folliculin is part of the autophagy signaling pathway to regulate epithelial cell size in response to fluid flow


FIGURE 1: Shear stress induces autophagy, cell size decrease and FLCN expression. (A-D) HK2 cells were subjected to fluid flow from 4 h to 4 days (shear 4 h, 48 h, 96h-4D), or not (static 4 h, 48 h, 96h-4D). (A) After fluid flow treatment or static culture, cells were fixed, labeled with DAPI and immunostained for the autophagosome marker LC3 or stained with phalloidin to reveal F-actin and cell boarders are marked out with white dashes (B) and then analyzed by fluorescence microscopy. (C) Quantification of LC3 puncta (LC3 dots number per 500 μm² area) from experiments shown in (A). (D) Quantification of cell area (mean) from experiments shown in (C). (E-F) HK2 cells were subjected to fluid flow from 4 h to 4 days (shear 4 h, 48 h, 96h-4D), or not (static 4 h, 48 h, 96h-4D). (E) Representative western blot analysis of FLCN, LC3I, LC3II and actin in the indicated conditions. (F) Quantification of Western blot shown in (E). (G) HK2 cells were subjected to 4 days (shear 4D) fluid flow or not (static 4D) and FLCN mRNA expression level was determined and quantified by RT-qPCR. (H) HK2 cells were maintained in normal culture condition (CTRL) or subjected to a 24 h serum starvation (SS 24 h). Levels of the FLCN, LC3I and LC3II were analyzed by western blot. Scale bars in (A) and (C) = 10μm.

By continuing to use the site, you agree to the use of cookies. more information

The cookie settings on this website are set to "allow cookies" to give you the best browsing experience possible. If you continue to use this website without changing your cookie settings or you click "Accept" below then you are consenting to this. Please refer to our "privacy statement" and our "terms of use" for further information.

Close