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. 2003 Sep;77(17):9685–9694. doi: 10.1128/JVI.77.17.9685-9694.2003

FIG. 6.

FIG. 6.

Northwestern analysis of VIRP1. Specificity of binding of VIRP1 protein with PSTVd positive-strand RNA was confirmed by Northwestern assay. Crude extracts of E. coli BL21 cells harboring either pHIS, pHis-VIRP1 or pHis1-VIRP1Δ or without plasmid (see the text for details) were separated by SDS-10% PAGE, transferred to nitrocellulose membranes and probed with radioactively labeled transcripts. In panels A and C, hybridization was done with PSTVd positive-strand RNA; in panel B, hybridization was done with U1 RNA. (A and B) BL21(pHIS) or BL21(pHIS-VIRP1) crude cell extracts. Lanes 1 and 3, noninduced cells; lanes 2 and 4, IPTG-induced cells. (C) BL21 cell extracts. Lane I1, BL21(pHis-VIRP1) cell extracts; lane I2, BL21 cell extracts; lane II1, BL21(pHis1-VIRP1Δ) cell extracts; lane II2, BL21 cell extracts. The arrows indicate the VIRP1 and VIRP1Δ proteins, respectively. In panel C, parts I and II present results from two different experiments.