Fig. 3.

Induced or exogenous expression of MAGEC2 is regulated by TRIM28. a Knockdown of TRIM28 decreases the expression level of MAGEC2 induced by demethylating agent 5-aza in A549 cells. TRIM28 specific siRNAs or control (si-NC) were transfected into A549 cells for 12 h, followed by adding 5-aza (10 μM) for 4 days, and cell lysates were immunoblotted with anti-MAGEC2 or anti-TRIM28 antibodies. Expression levels of GAPDH are indicated as an internal control. b Knockdown of TRIM28 decreases the exogenous expression level of MAGEC2 in Hs 695 T cells. FLAG-tagged MAGEC2 expression vector and TRIM28 specific siRNAs or si-NC were co-transfected into Hs 695 T cells for 48 h, and cell lysates were immunoblotted with indicated antibodies. In MAGEC2 blots, the upper band represents FLAG-tagged MAGEC2 and the lower band represents endogenously expressed MAGEC2. *, unspecific bands. c Overexpression of TRIM28 increases exogenous MAGEC2 protein level in AsPC1 cells. pCL-TRIM28 and FLAG-MAGEC2 or FLAG-MAGEA3 (as a control) expression vectors were co-transfected into AsPC1 cells for 48 h, and cell lysates were immunoblotted with indicated antibodies. Each experiment was repeated three times