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Fig. 1 | Cell Communication and Signaling

Fig. 1

From: DRAM1 regulates autophagy and cell proliferation via inhibition of the phosphoinositide 3-kinase-Akt-mTOR-ribosomal protein S6 pathway

Fig. 1

DRAM1 enhances autophagosome formation and downregulation of rpS6 phosphorylation. a HEK293T cells were transfected with FLAG empty vector or FLAG-DRAM1 for 24 h, the protein levels of p62, LC3, FLAG and β-actin were detected with Western blot analysis. b Quantitative analysis of the optical densities of LC3-II and p62 in HEK293T cells transfected with FLAG-DRAM1. Data represent mean ± SEM for combined data from three independent experiments. c HEK293T cells were transfected with FLAG empty vector or FLAG-DRAM1 for 48 h and then were treated with DMSO or Bafilomycin A1 (100 nM) for 6 h before harvesting. d Representative confocal images of HEK293T cells transfected with FLAG-DRAM1 or GFP-LC3 alone in control group and co-transfected with FLAG-DRAM1 and GFP-LC3. Scale bar represents 10 μM. e HEK293T cells were transfected with HA-Atg13, MYC-ULK1 and different amount of FLAG-DRAM1 for 24 h. The whole cell lysates (200 μg) were immunoprecipitated with an anti-IgG or anti-MYC antibody, and the precipitates were detected with an anti-HA antibody. f HEK293T cells were transfected with FLAG empty vector or FLAG-DRAM1 for 24 h, and the protein levels of p-rpS6 (S235/236), rpS6, p62, LC3, FLAG and β-actin were detected with Western blot analysis. Levels of p-rpS6 (S235/236) and rpS6 were quantitatively analyzed. Data represent mean ± SEM for combined data from three independent experiments. g Hela cells were transfected with FLAG empty vector or FLAG-DRAM1 for 36 h, and the protein levels of p-rpS6 (S235/236), rpS6, p62, LC3, FLAG and β-actin were detected with Western blot analysis. *p < 0.05 vs control

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