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. 2014 Apr 28;111(19):6970–6975. doi: 10.1073/pnas.1322545111

Fig. 2.

Fig. 2.

TRIM65 interacts and colocalizes with TNRC6. (A) HEK 293 cells stably expressing TRIM65-FLAG were treated with 10 μM protease inhibitor MG132 or DMSO for 4 h. Cell lysates were collected and treated with 100 μg/mL RNase A. Immunoprecipitation (IP) and immunoblotting were performed using the indicated antibodies. con, control; WB, Western blot. (B) GFP and various TRIM65 domains fused with GFP were transfected with TNRC6A-FLAG into HEK 293 cells. Cell lysates were coimmunoprecipitated with anti-FLAG antibody and blotted with the indicated reagents. The asterisk indicates GFP protein size. (C) HeLa cells were first treated with 10 μM MG132 or DMSO for 4 h and then fixed before incubation with anti-TNRC6A and anti-TRIM65 antibodies, followed by detection with secondary antibodies. DAPI was used to detect nuclei. Arrows highlight colocalization.