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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Cell Signal. 2014 Mar 20;26(7):1445–1455. doi: 10.1016/j.cellsig.2014.03.007

Fig. 5.

Fig. 5

Mono-ubiquitinated STAT3 induces expression of proliferation and anti-apoptotic marker gene expression. (A) HepG2 cells were transfected with either STAT3 or ubi-STAT3 FP. 40 h after transfection total RNA was isolated and Q-RT-PCR was performed for SOCS3, SOD2, APEX1, BCL2, BCL2L1expression. Data are expressed as mean ± SD, * indicates P< 0.05. (B) STAT3−/− MEF were transfected either with STAT3 or ubi-STAT3 FP and mRNA expression of Ref1, BCL2L1, CCND1, and SOD2 were measured by Q-RT-PCR. (C) HEK 293 cells were transfected with either STAT3 (S3) or ubi-STAT3 FP (ubi-S3). 24 h after transfection cells were treated with JQ1 (10 μM and 50 μM for overnight) and total RNA was isolated for mRNA expression of MYC. Data are expressed as mean±SD, * indicates P< 0.05. (D) STAT3−/− MEF were electroporated with either STAT3 or ubi-STAT3 FP. 40 h after transfection cells were treated with TNFα (15 ng/ml) and cyclohexamide (30μg/ml) for 0, 3 and 5 h. Cells were gated for mcherry positive cells and early apoptosis was measured by FITC conjugated Annexin V staining. Shown is the percentage of apoptotic cell death as measured by flow cytometry (FACS Canto, BD).