FIG 9.

CD163 is essential for PRRSV infection via macropinocytosis. (A and B) CD163 is required for PRRSV infection via macropinocytosis. BHK-21 cells were transfected with TIM-4 and/or CD163 for 36 h and then infected with PRRSV BJ-4 (MOI = 10) for 1 h. RT-qPCR and IB were performed with specific primers and antibodies at 12 and 24 hpi, respectively. (C) Colocalization of PRRSV and CD163 in SNX5-marked macropinosomes. PAMs were inoculated with an MOI of 10 PRRSV BJ-4 at 37°C for 30 min. Cells were fixed and stained with anti-SNX5 antibody and anti-TIM4 or anti-CD163 antibody. Nuclei were stained with DAPI, followed by confocal microscopy. The colocalization was assessed by determination of the Manders’ overlap coefficient. Each experiment was performed three times, and similar results were obtained. Scale bars, 10 μm.