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. 2021 Jul 28;13(8):1480. doi: 10.3390/v13081480

Figure 4.

Figure 4

Interaction between CD2v and CD58. (A,B) Interaction between CD2v and porcine CD58. (A) PK15 cells were cotransfected with plasmids pCD58-Flag and pCD2v-HA or pCD58-Flag and pEmpty-HA (control) and harvested at 8 h pt. Whole cell lysates (left) and extracts immunoprecipitated with anti-Flag antibodies or anti-HA were examined by Western blotting with antibodies directed against proteins indicated on the right. (B) PK15 cells were cotransfected with pCD58-Flag and pCD2v-HA, fixed at 24 h pt, incubated with mouse anti-Flag and rabbit anti-HA primary antibodies, washed and incubated with secondary antibodies (Alexa-fluor 488-labeled anti-mouse and Alexa-fluor 594-labeled anti-rabbit). Cells were counterstained with DAPI and examined with the confocal microscope. Insets show magnified areas of the field. (C) 293T cells were transfected with pCD2v-HA or pEmpty-HA (control) and harvested at 8 h pt. Whole cell lysates (left) and extracts immunoprecipitated with mouse anti-huCD58 antibodies or anti-HA were examined by Western blotting with antibodies directed against proteins indicated on the right. (D) 293T cells were transfected with pCD2v-HA, fixed at 24 h pt, incubated with mouse anti-huCD58 and rabbit anti-HA primary antibodies, washed and incubated with Alexa-fluor 488-labeled anti-mouse and Alexa-fluor 594-labeled anti-rabbit secondary antibodies. Cells were counter stained with DAPI and examined with the confocal microscope. Insets show magnified areas of the field. Results for (A–D) are representative of three independent experiments.