Figure 3: Effect of MYPT1 silencing on the methyltransferase activity of PRMT5 in HepG2 cells. | Scientific Reports

Figure 3: Effect of MYPT1 silencing on the methyltransferase activity of PRMT5 in HepG2 cells.

From: Myosin phosphatase and RhoA-activated kinase modulate arginine methylation by the regulation of protein arginine methyltransferase 5 in hepatocellular carcinoma cells

Figure 3

Immunofluorescent staining of non-target control (Ctrl) and MYPT1-silenced (siMYPT1) cells using antibodies specific for MYPT11-296 (A), PRMT5 (B), histone H2A symmetric dimethyl Arg3 (H2AR3me2s (C) and histone H4 symmetric dimethyl Arg3 (H4R3me2s (D) and actin as inducated in the figures. Scale bars: 50 μM. Enlargement of framed regions in merged images are shown on the right on A, B, C and D panels. Nuclear fractions of non-target control (Ctrl) and MYPT1-silenced (siMYPT1) HepG2 cells were prepared and analysed by Western blot using anti-MYPT11-296 (E), anti-PRMT5 (F), anti-pPRMT5T80 (G), anti-histone H2A symmetric dimethyl Arg3 (H) and anti-histone H4 symmetric dimethyl Arg3 (I) specific antibodies. The relative expression of MYPT1 or PRMT5 and the relative symmetric dimethylation level of H2AR3 or H4R3 were normalized to lamin A/C as internal control. Samples derived from the same experiment and the blots were processed in parallel or assayed after stripping. The relative phosphorylation level of PRMT5T80 was normalized to the expression level of PRMT5 and then to lamin A/C as internal control by densitometry. Mean ± SEM; n = 3; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by student t-test.

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