Figure 1.

Inhibiting EV-A71 Replication by Knockdown of Pim1

(A) RD cells were infected with EV-A71 or inactive EV-A71 (UV light-inactive virus, negative control) at MOI of 10, and cellular mRNA was extracted at different time points p.i. Pim1 mRNA level was determined by qRT-PCR.

(B and D) Pim1 was silenced by two individual siRNA duplexes si-Pim1-1 and si-Pim1-2. The relative cellular Pim1 mRNA level was quantified by qRT-PCR at 48 h after transfection in both (B) RD and (D) HeLa cells. GAPDH was used as the internal control.

(C and E) (C) RD and (E) HeLa cells treated with Pim1-specific siRNA and infected with EV-A71 at MOI 0.01. The extracellular viral RNA level was calculated at 48 h.

(F and G) Viral titer was further determined in both (F) RD and (G) HeLa cells.

(H) Cell survival image was taken in RD cells.

(I and J) Pim1 was silenced in EV-A71-infected (J) RD and (I) HeLa cells.

(K) The quantification results of (I).

(L) The quantification results of (J). Protein levels of viral VP1 protein were detected and quantitated. The density of VP1 to β-actin was set as 1, and the relative fold change is indicated.

Data are represented as mean ± SD (n = 3). Student's t test, *p < 0.05, compared with mock group; **p < 0.01, compared with mock group.