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. 2022 May 21;23(10):5773. doi: 10.3390/ijms23105773

Figure 2.

Figure 2

Characterisation of UL34 as an augmenting viral protein with leaky late expression kinetics. (A) Intracellular HCMV genome copies at 120 HPI, relative to genome copies at 12 HPI. MOI = 3, n = 3, bars = SD. (B) Western blot analysis of UL34 expression in MRC5 cells treated with 100 μg/mL phosphonoacetic acid (PAA) and subsequently infected with either AD169-GFP WT or AD169 HA-UL34 HCMV (5 DPI, MOI = 3). Membranes were probed with primary antibodies against HCMV viral proteins, HA, or β-actin loading control. (C) Immuno-fluorescence analysis of host GM130 and viral UL99 in WT MRC5 cells infected with WT or ΔUL34 AD169-GFP virus. 4 DPI, MOI = 0.1, scale bars = 20 μm. (D) Growth kinetics of ΔUL34 AD169-GFP virus, as measured by IE1 fluorescent focus assay in cell culture supernatants from WT and UL34-complementing MRC5 cells. MOI = 3, n = 3, bars = SD. (E) Spread of ΔUL34 AD169-GFP virus in WT and UL34-complementing MRC5 cells, as quantified by fixing, staining, and counting IE1 positive cells at indicated time points. MOI = 0.01, n = 3, bars = SD.