V protein interacts with TXNL1 and downregulates the expression of TXNL1. A Interaction between V and TXNL1 in the yeast system. The white clones show that the bait (pBD-V) had no autoactivator activity in yeast. When pGBKT7-V and pGADT7-TXNL1 were cotransformed into the Y2H Gold yeast strain, the cotransformants turned blue on DDO/X/A and QDO/X/A plates after incubation. At the same time, the pGBKT7-p53 and pGADT7-T cotransformants were used as positive controls, whereas the pGBKT7-Lam and pGADT7-T cotransformants were used as NC. B Confocal analysis of TXNL1 and V in DF-1 cells. V protein was transfected into DF-1 cells and assessed by immunofluorescence staining. V protein was detected with goat antimouse monoclonal antibody (MAb) and visualized with Alexa Fluor 488 (green). TXNL1 was detected with a rabbit anti-TXNL1 polyclonal antibody and visualized with Alexa Fluor 546 (red). Yellow indicates colocalization of the V protein and TXNL1 in the merged image C V protein downregulated the expression of TXNL1. DF-1 cells were transfected with V and control plasmids (pCAGEN) for 48 h. The RNA was extracted according to the method previously described. The mRNA levels of endogenous TXNL1 were detected using Q-PCR. D DF-1 cells were transfected with V and control plasmids (pCAGEN) for 48 h. Whole-cell extracts were prepared for Western blot analysis that was specific for the indicated proteins.