Fig. 8
From: Autophagy acts through TRAF3 and RELB to regulate gene expression via antagonism of SMAD proteins
Inhibition of SMAD4 rescues the effects of autophagy loss in vivo. a A549 WT and ΔATG5 cells were transduced with control non-targeting shRNA (shGFP) or shRNA targeting SMAD4 (shSMAD4). Stable lines were immunoblotted as indicated. b Cells were plated for time-lapse phase-contrast videomicroscopy using an Incucyte microscope and cell proliferation was monitored by automated confluency analysis at set intervals post plating (n = 9 wells, ± S.D.). c Cells were subcutaneously injected into immunocompromised mice and tumour volume was monitored longitudinally (means, n = 10 flanks, ±S.E.M., *P < 0.05 vs. WT shGFP, two-tailed t-test). d, e A549-Cas9 and cognate A549 ΔFIP200 cell lines were transduced with shGFP or shSMAD4. Selected pools were d immunoblotted as indicated or e subcutaneously injected into immunocompromised mice, whereupon tumour volume was monitored longitudinally (means, n = 8 flanks, ±S.E.M., *P < 0.05 vs. all other conditions, two-tailed t-test). Uncropped blots are available in Supplementary Fig. 10
