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. 2021 Dec 23;12:795730. doi: 10.3389/fmicb.2021.795730

FIGURE 6.

FIGURE 6

Effect of overexpression of pUL48 and its truncated protein on viral IE gene transcription. The relative transcription level of the IE gene at 4/8/12 h after infection with MOI = 1 DPV-CHv; pCAGGS, pCAGGS-pUL48-HA, pCAGGS-pUL48 (Δ1–20 aa)-HA, and pCAGGS-pUL48 (Δ1–60 aa)-HA were transfected into a 12-well plate DEF, respectively. At 12 h after transfection, DEF was infected with DPV CHv strain with MOI = 1. The virally infected cells were collected 4, 8, and 12 h after infection into the non-RNA enzyme EP tube using RNAiso Plus. According to the instructions of the RNA extraction kit, reverse transcription reagent and SYBR®Premix Ex Taq™ II Kit, RNA extraction, reverse transcription, and quantitative fluorescence PCR were performed successively to detect the mRNA levels of IE gene ICP4, ICP22, and ICP27. The expression of each truncated protein of pUL48 was detected by WB. 1, pCAGGS; 2, pUL48; 3, pUL48 (Δ1–20 aa); 4: pUL48 (Δ1–60 aa). Relative quantitative test results were processed using 2– ΔΔCt, and the results were normalized to the control group. Student’s t-test was used to analyze the experimental and control groups differences. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.