Fig. 3

Effects of autophagy induced by MHBSt¹⁶⁷ on cell proliferation and the cell cycle in L02 cells. A Western blotting analysis of autophagy-related proteins (LC3B and p62) and statistical analysis. Cells were treated with DMSO, 100 nM rapamycin (Rapa), and 2 mM 3-MA for 2 h and then transfected with MHBS and MHBSt¹⁶⁷ expression plasmids for 48 h. The individual gray value in Western blots was measured and normalized against β-actin, then the relative intensity of target protein to β-actin was calculated by setting the control vector transfection as 1.00, *P < 0.05. B Cell viability was measured by CCK-8 assays in L02 cells. Before transfection with the MHBSt¹⁶⁷ expression plasmid, cells were treated with DMSO, 100 nM Rapa, and 2 mM 3-MA for 2 h. The results are expressed as the OD values from three experiments performed in duplicate. *P < 0.05. C Cell proliferation was measured by high-content screening (HCS) assays. The results are expressed as cell count values obtained every 0.5 h for 72 h. D–F Cell cycle analysis was performed by flow cytometry. Cells were harvested after being transfected with plasmids for 48 h and stained with PI. Representative images from three independent experiments are shown