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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

Fig. 4

RPS27a directly binds RPL11, and the knockdown of RPS27a regulates p53 activation. A Docked positions of RPS27a and RPL11, shown as a cartoon model in light green and yellow. The ligands are represented as sticks in magenta, cyan and yellow-green. B Binding analysis of RPS27a and RPL11 in vitro with GST pull-down assays. Fusion protein beads were used for pull-down and detected with IB with anti-GST and anti-S-Tag antibodies; Coomassie staining of GST and GST-RPS27a proteins is shown in the upper panel; IB analysis is shown in the middle and lower panels. C A549 cells were transfected with Flag-RPS27a plasmids and harvested for co-IP assays with anti-Flag antibody. D A549 cells were transfected with RPS27a-siRNA for 48 h and treated with 40 μM MG132 for 4 h. The cell lysates were subjected to IP with anti-RPL11 antibody, followed by IB with antibodies to MDM2, RPS27a and RPL11. E A549 cells were transfected with His-Ub plasmids and RPS27a-siRNA, then treated with 40 μM MG132 for 4 h. The cell lysateswere subjected to IP with anti-His antibody, followed by IB with anti-p53 antibody to detect ubiquitinated p53. NC, negative control; IP, immunoprecipitation; co-IP, co-immunoprecipitation; IB, immunoblotting

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