FIG 9.

RPLP1/2 knockdown impairs accumulation of DENV-2 structural proteins expressed in stable cell lines. Tetracycline-inducible HeLa and HEK-293 cells expressing C-prM-E were transfected with the indicated siRNAs, and 48 h later tetracycline was added to the medium for 24 h. Cell lysates were harvested for Western blot analysis 24 h after addition of tetracycline. (A) Representative Western blots showing expression of C-prM and E proteins under NSC or siP2_4 transfection conditions (performed in triplicate). The arrows indicate cleaved E protein, the 37-kDa C-prM minority species, and the 33-kDa C-prM majority species. Samples from uninduced cells are also indicated (-tet). (B) Quantifications of C-prM-E RNA by RT-qPCR normalized to 18S rRNA and protein bands in the Western blot assay from HeLa cells, normalized to results with β-actin. The NSC conditions were set to 100%. Graphs show means and standard deviations from quantification of two independent assays performed in triplicate. (C) The same experiment in shown in panel B, except HEK-293 cells were used. Graphs show means from the quantification of three independent assay results. Statistical significance was assessed by a two-tailed Student's t test between NSC and experimental siRNA conditions. *, P < 0.05; ****, P < 0.0001.