Fig. 5

LukS-PV increasing α-tubulin acetylation through HDAC6 downregulation. A A whole-cell extract from Huh-7 cells stably expressing HDAC6 was subjected to affinity purification with protein A/G beads. The purified protein complex was resolved by SDS–polyacrylamide gel electrophoresis followed by staining with Coomassie brilliant blue. The bands were retrieved and analyzed by mass spectrometry. The peptide (AVCMLSNTTAIAEAWAR) showing in this panel is the α-tubulin sequence detected by mass spectrometry. B The whole-cell lysate and sample from the co-immunoprecipitation with anti-HDAC6 antibody in HCC cells were verified by western blot. C-D The indicated proteins were verified by western blot after transfection of HCC cells with the HDAC6 plasmid. Relative band intensities (compared to GAPDH) were measured using Image J. E–F The expression levels of α-tubulin and its acetylated form were determined in HCC cells after treatment with different concentrations of LukS-PV. Relative band intensities (compared to GAPDH) were measured using Image J. * P < 0.05, ** P < 0.01, or *** P < 0.001