Fig. 6

Effects of MHBSt¹⁶⁷-induced autophagy on NF-κB activation in L02 cells. A Western blotting analysis of NF-κB activation and statistical analysis. Before transfection with MHBS and MHBSt¹⁶⁷ expression plasmids, cells were treated with DMSO, 100 nM Rapa and 2 mM 3-MA for 2 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–D Confocal microscopic analysis of NF-κB activation. Before transfection with MHBS and MHBSt¹⁶⁷ expression plasmids, cells were treated with DMSO, 100 nM Rapa and 2 mM 3-MA for 2 h. The intracellular localization of p-NF-κB/p65 was determined by immunofluorescence staining using an anti-p-NF-κB/p65 antibody. Nuclei were stained with DAPI. Scale bar: 50 μm. E Analysis of red fluorescence intensity in nuclear in B, C, D. Representative images from three independent experiments are shown