Fig. 7

HDAC6 silencing attenuated lung cancer growth through a GRP78-ERK-dependent mechanism. A H460 and B A549 cells were transfected with siHAC6, siGRP78, or control siRNA (siCtrl). Cell proliferation was examined by an MTT assay. Data are presented as the mean ± SEM. *p < 0.05 vs. initial time point (0 h); ^#p < 0.05 vs. siCtrl group (n = 3). Patient-derived lung cancer C ELC12 and D ELC16 cells were transfected siHDAC6, siGRP78, or control siRNA (siCtrl), and seeded onto ultralow attachment plates for allowing 3D tumor spheroid formation. The tumor spheroid growth rate was quantified from the spheroid diameter using ImageJ software and calculated relative to that of the initial time point. Data are presented as the mean ± SEM. *p < 0.05 vs. initial time point (0 h); ^#p < 0.05 vs. siCtrl group (n = 3). Scale bar is 100 µm. After 5 d of inoculation, tumor spheroids growth from transfectant C ELC12 and D ELC16 cells were lysed. The expression levels of GRP78, p-ERK, ERK, and acetylated tubulin were analyzed by immunoblotting. The intensity was normalized to that of GAPDH. Data are presented as the mean ± SEM. *p < 0.05 vs. siCtrl cells; ^#p < 0.05 vs. between indicated groups (n = 3)