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. 2015 Sep 25;6(35):37706–37723. doi: 10.18632/oncotarget.5342

Figure 2. MAT2A and sumoylation machinery regulate Bcl-2 expression in HepG2 cells.

Figure 2

A. HepG2 cells were transfected with siMAT2A, siUbc9 and siSUMO-1 (20 nM) or scrambled siRNA (Sc) for 48 hours and/or MAT2A overexpression vector or empty vector (EV) for 24 hours. The mRNA levels of Bcl-2 were compared to Sc+EV using real-time PCR. Results represent mean ± SEM from 4 experiments in duplicates, *p < 0.04 vs. Sc+EV control; †p < 0.04 vs. Sc+MAT2A overexpression vector. B. Following the same treatments as in “A”, HepG2 cells were transfected with the human Bcl-2 promoter or pGL3-basic as described in Methods. The luciferase activity driven by Bcl-2 promoter was normalized to that of pGL3-Basic and expressed as % over Sc+EV. Results represent mean ± SEM from 4 experiments in duplicate, *p < 0.04 vs. Sc+EV control; †p < 0.03 vs. Sc+MAT2A overexpression vector. C. Cells were treated as in ‘A’ and total cellular protein was subjected to Western blotting with antibodies against MATα2 or Bcl-2. Densitometric changes were normalized to actin. Representative images and densitometric analysis (% mean ± SEM of Sc+EV, indicated below the blots) from 3 experiments in HepG2 cells are shown. *p < 0.05 vs. Sc+EV, †p < 0.03 vs. Sc+MAT2A overexpression vector.