Figure 5

Nuclear translocation of TFEB is important for PDCoV-induced lipophagy. A–C LLC-PK1 cells were infected with PDCoV (MOI = 0.1) and treated with 50 μM 25HC for indicated time. A Cells were imaged by confocal microscopy using TFEB-specific antibody, TFEB was stained green and nuclei were stained blue with DAPI. Scale bar, 20 μm. Mean fluorescence intensity of TFEB were analyzed by Image J. B cytoplasm and nuclei extracts underwent western blot with TFEB-specific antibody. Histone-H3 and GAPDH were used as nuclear control and cytoplasmic control, respectively. C Immunoblot bands in panel B were quantified by Image J. D LLC-PK1 cells were transfected with four siRNAs targeting TFEB, the relative mRNA expression of TFEB was analyzed by qPCR. E–J LLC-PK1 cells were transfected with the second siRNA in panel D, followed by infection with PDCoV for 12 h. E andG The indicated protein expressions were analyzed by western blot, GAPDH was used as a loading control. F and H Bands in panel E and G were quantified by Image J. I and J Relative mRNA expressions of indicated genes were analyzed by qPCR. Error bars, mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.