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. 2024 May 30;12:RP90316. doi: 10.7554/eLife.90316

Figure 3. Nonstructural protein 5 (Nsp5) expression induces cleavage of tRNA methyltransferase 1 (TRMT1) in human cells.

Figure 3.

(A) Immunoblot of lysates prepared from human 293T cells expressing GFP, Nsp5 or Nsp5-C145A. The immunoblot was probed with anti-TRMT1, Strep, or actin antibodies. Hours post represents the time post-transfection. Circle represents endogenous TRMT1. Arrow represents the N-terminal (N)-TRMT1 cleavage fragment. Asterisk (*) denotes a non-specific band. Size markers to the left in kiloDalton. (B, C) Quantification of endogenous TRMT1 or N-terminal (N)-TRMT1 cleavage product in transfected cells. TRMT1 and N-TRMT1 signal was normalized to actin. (D) Immunoblot of lysates prepared from wild-type or TRMT1-knockout (KO) human cell lines expressing Nsp5. Experiments in (A) and (D) were repeated three times in biological replicates (see source data).

Figure 3—source data 1. Raw uncropped immunoblots for Figure 3.