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. 2011 Dec 23;287(10):7313–7323. doi: 10.1074/jbc.M111.331660

FIGURE 3.

FIGURE 3.

Impact of PRMT5 on PKCδ-dependent hINV promoter activity. A, KERn were electroporated with 3 μg of control- or PRMT5-pooled (Santa Cruz) siRNA, and after 48 h the cells were trypsinized and electroporated with 1.5 μg of pINV-2473 (involucrin promoter luciferase reporter) in the presence of 1.5 μg of empty vector (pEGFN1) or PKCδ-encoding plasmid (pPKCδ-EGFN1). After an additional 18 h, cell extract was prepared for assay of luciferase activity. Values are the mean ± S.E., n = 3. The control-siRNA EV and PKCδ values are significantly different, p < 0.001; the PRMT5-siRNA EV and PKCδ values are significantly different, p < 0.001; the Control-siRNA PKCδ and PRMT5-siRNA PKCδ values are significantly different, p < 0.001. B, KERn were electroporated with 1.5 μg of pcDNA3 or pcDNA3-PRMT5 expression vector, and after 48 h cells were re-electroporated with 1.5 μg of pINV-2473 and 0.5 μg of pEGFN1 or pPKCδ-EGFN1. After an additional 18 h, extracts were prepared for luciferase assay. The values are the mean ± S.E., n = 3. The pcDNA3 EV and PKCδ values are significantly different, p < 0.001. C, KERn were incubated with the indicated expression vectors or siRNA, and after 24 h extracts were prepared for detection of the indicated proteins by immunoblot. Similar results were observed at 48 and 72 h (not shown).