Figure 6
HSV-1 and CMV infection-induced changes in the human TM cell actin cytoskeletal organization, and accumulation of viral DNA at 12 h and 2 days post-infection. Primary TM cells obtained from ScienCell Research Labs were infected with HSV-1 or CMV alone, or with HSV-1 and CMV at a multiplicity of infection 1, and observed at 12 h (A,C,E,G) and 2 days post-infection (PI) (B,D,F, H). Stress fibers were stained with a Rhodamine Phalloidin (red signals) (A–H). Increased contraction of F-actin was observed after HSV-1 infection and CMV infection (C–F) when compared with mock infection (A,B). Moreover, the co-infection of HSV-1 and CMV also caused increase in F-actin formation (G,H). Cells were harvested and viral DNA was extracted from cells and processed for qPCR analysis of viral DNA accumulation using HSV-1 DNA polymerase and UL26 primer for HSV-1 (I) and CMV (J), respectively. Real-time PCR with β-actin primers were performed to serve as an internal control for input DNA. Data are the averages of three independent DNA samples from the infected cells. Activation of RhoA activity in TM cells at 2 days PI was significant after co-infection of CMV and HSV-1 in immunoblotting (K) and densitometric analyses (L). The immunoblot data were normalized to total Rho. Blots were cut prior to hybridization with antibodies. Data are shown as means ± SEM, N = 3. *P < 0.05 and **P < 0.01 vs. transcripts from the mock infection within 12 h and 2 day PI using one way ANOVA with Dunnett’s multiple comparison.
