Figure 5: C-terminus of MYH9 is the binding domain of PRRSV GP5.
From: MYH9 is an Essential Factor for Porcine Reproductive and Respiratory Syndrome Virus Infection
(a) PK-15-NFlag, PK-15-GP5Flag, COS-7-NFlag, and COS-7-GP5Flag cells were incubated with purified PRAHis at 4 °C for 1 hr, fixed after shifting to 37 °C for 30 min and imaged by confocal microscopy after stained with antibodies specific for His (green) and FLAG (red). Nuclei (blue) were counterstained with DAPI. Scale bars, 5 μm. (b) Anti-PRAHis serum dilutions were mixed with PRRSV-EGFP. The mixtures were incubated with MARC-145 cells at 4 °C for 1 hr, then cells were harvested and relative PRRSV binding levels were analyzed by quantitative RT-PCR. (c) After incubating with the mixtures at 4 °C for 1 hr, the cells were shifted to 37 °C for 30 min, and then washed to remove unbound virus. The quantity of PRRSV positive infected cells was analyzed by FACS 48 hrs post infection. Pre-immunization serum (pre) at 1:10 served as the control. (d) After cells were incubated for 1 hr at 4 °C in the mixtures, the cells were washed with DMEM prior to a shift to 37 °C for 48 hrs. The number of PRRSV positive infected cells was analyzed by FACS. Pre-immunization serum (pre) at 1:10 served as the control. In both (c) and (d) the data were shown as relative percentages compared to the value of the pre-immunization serum sample, whose mean value was set as 100% infection. Error bars, mean ± s.e.m. (n = 3). *P < 0.05.
