FIG 5.
RBM38 regulates 11-kDa protein expression. CD36+ EPCs transduced with scramble shRNA (shScram)- and shRBM38-1 and -3 (shRBM38)-expressing lentiviruses were infected with B19V at 2 days postransduction. UT7/Epo-S1 and UT7/Epo-S1RBM38KD cells were transfected with M20. (A) Western blotting. The cells were collected at 2 days postinfection or posttransfection, lysed, and run for Western blotting. Immunoblots were probed for the NS1, VP1/2, and 11-kDa proteins, using their respective antibodies. Blots were reprobed for GAPDH as a loading control. (B) RT-qPCR. Specific primers and probes for the detection of mRNA that encodes the VP2 or 11-kDa protein, as diagrammed, were used for qPCR analysis. (C to F) Detection of mRNAs that encode the 11-kDa, VP2, NS1, and VP1 proteins by RT-qPCR. At 2 days posttransfection, total RNA was extracted from UT7/Epo-S1 and UT7/Epo-S1RBM38KD cells. The RNA was further used to generate cDNA for the quantification of mRNAs that encode the 11-kDa (C), VP2 (D), NS1 (E), and VP1 (F) proteins, using the RT-qPCR approach. Quantified viral mRNA levels were normalized to the level of β-actin mRNA, and mRNAs extracted from M20-transfected control UT7/Epo-S1 cells were used as controls and arbitrarily set as a value of 1. n.s, no statistical significance (P > 0.05); ****, P < 0.0001.