Fig. 4: MAT1A inhibited the ubiquitination of CCND1 by blocking the binding of SKP2 to CCND1.
From: MAT1A activation of glycolysis to promote NSCLC progression depends on stabilizing CCND1
In co-immunoprecipitation experiments, the total lysates of A549 and NCI-H1299 cells were immunized with (A) anti-CCND1 or (B) anti-SKP2 and WB, respectively. In the co-immunoprecipitation experiments, total lysates of A549 and NCI-H1299 cells were immunized with (C) anti-MAT1A or (D) anti-SKP2 and WB. E MAT1A knockdown and control A549 and NCI-H1299 cells were treated with protein synthesis inhibitor (CHX, cycloheximide from microbial, 10 μmol/L) and then subjected to cycloheximide chase assays. F SKP2 overexpressed and control A549 and NCI-H1299 cells were treated with CHX (10 μmol/L) to detect the relative protein levels of CCND1 at indicated time points. CCND1 protein level was determined in (G) MAT1A-depleted or (H) SKP2-overexpresed A549 and NCI-H1299 cells treated with or without proteasome inhibitor (MG132, 10 μmol/L). The ubiquitin of CCND1 immunoprecipitated in (I) MAT1A-depleted or (J) SKP2-overexpresed A549 and NCI-H1299 cells was detected. K Total lysates of A549 and NCI-H1299 cells with or without MAT1A knockdown were immunized with anti-CCND1 and WB.
