Figure 4: Caveolin-1 interact with TFV MCP. | Scientific Reports

Figure 4: Caveolin-1 interact with TFV MCP.

From: Caveolae Restrict Tiger Frog Virus Release in HepG2 cells and Caveolae-Associated Proteins Incorporated into Virus Particles

Figure 4

(A) CO-IP assay. HeLa cells were transiently transfected with myc-tagged TFV MCP and pCMV-flag empty vector (lanes 1 and 2), Flag-tagged caveolin-1 protein and myc-tagged TFV MCP, (lanes 3 and 4), pCMV-myc empty vector and FLAG-tagged caveolin-1 protein (lanes 5 and 6), respectively, as indicated (lane 3). Immunoprecipitation (IP) of Myc-tagged TFV MCP with Myc tag-specific monoclonal antibody led to coprecipitation of Flag-tagged caveolin-1 protein. IP of caveolin-1 protein Flag-tagged using Flag-tag-specific monoclonal resulted in coprecipitation of Myc-tagged TFV MCP. (B) HepG2 cells were infected with TFV at an MOI of 10. At 72 h postinfection, we collected the cells to do the immunoprecipitation using anti-caveolin-1 antibody (TFV infected), uninfected HepG2 cells as control (con). (C) Colocalization of caveolin-1 with TFV MCP was examined in HeLa cells transiently expressing Flag-tagged caveolin-1 protein and Myc-tagged TFV MCP. The intracellular localization of TFV MCP and caveolin-1 was analyzed by IFA using anti-Flag and anti-Myc antibodies. Caveolin-1 (red fluorescence) and TFV MCP (green fluorescencce) were viewed under a confocal microscope equipped with 555/488 nm argon-krypton and 543 nm helium-neon lasers. The yellow overlay represents colocalization of TFV MCP and caveolin-1. All nuclei were stained with a blue fluorescent dye (Hoechst 33342).

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