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

FIGURE 2.

FIGURE 2.

PRMT5 regulates hINV mRNA and protein level. A, KERns were electroporated with 3 μg of control, PRMT5-pooled (Santa Cruz), PRMT5-1, or PRMT5-2 siRNA (35), and after 48 h, PRMT5 and hINV protein levels were assessed by immunoblot. B, KERns were electroporated with 3 μg of the control (C), PRMT5-pooled (P, Santa Cruz), or individual (PRMT5-1, 1) or PRMT5-2, 2) siRNA and after 48 h PRMT5 and hINV mRNA was measured, by qPCR. Bars indicate relative levels of PRMT5 and hINV RNA normalized relative to GAPDH RNA levels and are the mean ± S.D. for triplicate samples in a single experiment. Similar results were obtained in three independent experiments. C, KERns were electroporated with 3 μg of control or PRMT5-pooled (Santa Cruz) siRNA. hINV, PRMT5, and SDMA were measured in total cell extracts prepared at 48 h post-electroporation. The arrows in the SDMA blot indicate the major SDMA-modified proteins. β-Actin is the control to normalize protein loading. D and E, KERn were electroporated with 3 μg of pcDNA3 or pcDNA3-hPRMT5, PRMT5 and hINV mRNA levels were measured after 24 h, and protein levels were measured at 48 h post-exposure. The values are the mean ± S.D. of triplicate samples in a single experiment. Similar results were observed in each of three experiments. The arrows in the SDMA blot indicate the major SDMA-modified proteins.