Figure 5: Production of TFV was inhibited by RNA interference of Nedd4.1. | Scientific Reports

Figure 5: Production of TFV was inhibited by RNA interference of Nedd4.1.

From: Budding of Tiger Frog Virus (an Iridovirus) from HepG2 Cells via Three Ways Recruits the ESCRT Pathway

Figure 5

(A) 100 nM siRNA specific for Nedd4.1/Nedd4.2 and control RNA (siGFP) were transfected into cells. The cells were collected from 24 h to 72 h and then subjected to Western blot. (B,C) 100 nM siNedd4.1/Nedd4.2 or 100 nM siGFP were transfected into cells 6 h prior to infection with TFV for 1 h and then cultured for 66 h. Cells and supernatant were lysed to extract DNA. Samples were then subjected to qPCR using MCP primers to quantify TFV genomic concentration. Statistical analysis was performed using Student’s t-test (0.05 < *p ≤ 0.1, 0.01 < **p ≤ 0.05, 0.01 ≤ ***p). (D) In vivo confocal microscopy analysis was performed after HepG2 cells were infected with TFV for 1 h and cultured for 66 h. Cells were processed for immunostaining as described under “Materials and Methods”. Arrows indicate co-localization of TFV MCP and endogenous Nedd4.

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