Fig. 4

Knockdown of LRRC15 promotes translocation of p65 from cytoplasm to nucleus. a RT-qPCR results showing expression of p65 target genes was enhanced when LRRC15 was silenced with a short hairpin RNA. b Protein levels of p-IκBα and total IκBα measured by western blot and quantitative analysis in the absence and presence of TNF-α for 30 min. c Nuclear and cytoplasmic proteins levels of p65, tubulin, and PCAF measured by western blot analysis after subcellular fractionation in LRRC15sh cells untreated or treated with TNF-α for 30 min. Cellular localization of endogenous p65 also observed by confocal microscopy in both control and LRRC15 knockdown cells with or without TNF-α treatment. Scale bars 100 μm. d Nuclear and cytoplasmic proteins levels of p65, tubulin, and PCAF were measured by western blot analysis after subcellular fractionation in LRRC15-overexpresssed cells untreated or treated with TNF-α for 30 min. Cellular localization of endogenous p65 also observed by confocal microscopy in both control and LRRC15-overexpressed cells with or without TNF-α treatment. Scale bars 50 μm. All data shown as mean ± SD, n = 3. **P < 0.01. NC negative control cells, LRRC15sh LRRC15 knockdown cells, LRRC15 fifteen-leucine-rich repeat containing membrane protein, IL interleukin, TNFα tumor necrosis factor alpha, ICAM intracellular adhesion molecule, GAPDH glyceraldehyde-3-phosphate dehydrogenase, DAPI 4',6-diamidino-2-phenylindole