FIG 1.

The RPLP1/2 heterodimer and RPLP0 are required for efficient DENV-2 and YFV infection of A549 and HuH-7 cells. Cells were transfected with either a nonsilencing control siRNA (NSC) or one of five independent siRNAs used to deplete RPLP1/2, three targeting RPLP1 (siP1_1, siP1_2, and siP1_6) and two targeting RPLP2 (siP2_1 and siP2_4). After 48 h, cells were infected at an MOI of 1 and infection was assessed after 24 h. (A and B) Western blotting results show knockdown of RPLP1/2 with independent siRNAs in A549 (A) and HuH-7 (B) cells. (C) Representative images showing A549 cells infected with DENV-2 (New Guinea C). Nuclei were Hoechst stained (blue), and the viral E protein was stained with 4G2 antibody (green). (D and E) Quantification of infection rates for DENV-2 and YFV (17D) are shown for A549 (D) and HuH-7 (E) cells. (F) A549 cells were transfected with the indicated siRNAs against RPLP0 and infected with YFV 48 h later at an MOI of 1. Western blotting results show knockdown of RPLP0 with two independent siRNAs. (G) Rates of infection are shown for cells transfected with NSC or siRNAs targeting RPLP0. The error bars represent standard deviations of three biological replicates. Statistical significance was assessed by a two-tailed Student's t test between NSC and experimental siRNAs. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.