Figure 5.

Inhibiting 2A^pro-mediated eIF4G Cleavage by CX-6258

(A) RD cells were treated with different concentrations of CX-6258 and then infected with EV-A71 at MOI 10. After culture for 9 h, cell lysates were collected and the cleavage of eIF4G was determined.

(B) The quantification results of (A).

(C) HEK293T cells were transfected with p2A^pro (0.1, 0.5, and 1 μg) for 36 h, and the cleavage of eIF4G was determined.

(D) HEK293T cells were first treated with different concentrations of CX-6258 for 2 h and transfected with 1 μg 2A^pro-expressing plasmid for 36 h. The cleavage of eIF4G was determined.

(E and F) The quantification results of (C) and (D), respectively.

(G) 293T cells were transfected with a Pim1-expressing plasmid for 48 h and then infected with EV-A71 at MOI 10 for the indicated time. The cleavage of eIF4G was determined.

(H) The quantification results of (G).

(I) 293T cells were first transfected with Pim1-specific siRNA for 48 h and then treated with CX-6258 at the indicated concentrations for 2 h, followed by transfection of a 2A^pro-expressing plasmid at 48 h. Cell lysates were collected, and the status of eIF4G cleavage was examined.

(J) The quantification results of (I). Protein levels were detected and quantitated by image density. The density of individual protein to β-actin or GAPDH in the control was set as 1, and the relative fold change is indicated.

Results are represented as mean ± SD from three independent experiments. *p < 0.05, **p < 0.01.