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. 2020 Feb 5;12(3):2507–2529. doi: 10.18632/aging.102760

Figure 5.

Figure 5

TRIM21 interacts with PRMT5 in U2 OS cells. (A) Colocalization of TRIM21 and PRMT5 was observed in U2 OS cells using antibodies against TRIM21 (green) and PRMT5 (red). Scale bar = 20 μm. (B) DAPI was used to indicate nuclei. Myc-VN155-PRMT5 and HA-VC155-TRIM21, along with HA-cerulean, were cotransfected into U2 OS cells for 48 h, and the reconstituted Venus fluorophore (yellow, arrows) was visualized via confocal microscopy. Scale bar = 20 μm. (C) The endogenous interaction between TRIM21 and PRMT5 was validated using a co-IP assay. (D, E) shRNAs targeting TRIM21 (shT#1 and shT#2) or plasmid encoding HA-TRIM21 were applied to knock down or overexpress TRIM21, and the protein and mRNA levels of TRIM21 and TXNIP were then determined by WB or quantitative real-time PCR, respectively. (F) Dox-inducible TRIM21-expressing cells was treated with MG132 (10 μM) for 12h, the protein expression of TRIM21, TXNIP, and p21 was then determined by WB.