Figure 3.

Knockdown of RACK1 abrogated PRRSV replication and NF-κB activation in Marc-145 cells. RACK1 mRNA expression level was measured by RT-qPCR at different time points post siRNA knockdown (A). The data was normalized to scramble siRNA treatment and the mRNA expression of internal control GAPDH. Forty-eight hours post siRNA knockdown, Marc-145 cells were inoculated with PRRSV YN-1 strain (25 TCID₅₀). Total protein from various infection time points was analyzed by western blot for RACK1 (B), viral N protein (C), p-IκBα (D), p65 (E), p-p65 (F) and GAPDH (G). siRACK1_1 and siRACK1_2 are the two siRNA sequences (see Table 1) used in this study targeting different RACK1 mRNA sequences. After SDS-PAGE and protein transferring, the membranes were cut based on the molecular weights of the target proteins and the markers, put together along the same film for exposure with same exposure time for each sub-figure. The histograms and blots shown here are representative data from three independent experiments.