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. 2018 Feb 5;217(2):763–777. doi: 10.1083/jcb.201705031

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© 2018 Viotti et al.

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Figure 6. — Overexpression of SUV420H2 in epithelial pancreatic cancer cells. (A) qRT-PCR for SUV420H2 in two epithelial pancreatic cancer cell lines transfected with an overexpression plasmid containing an SUV420H2 sequence with a deletion in the methyltransferase region (MT Dead) or a wild-type sequence. The qRT-PCR region probed is upstream of the deletion, near the 5′ end of SUV420H2. Data are normalized to HCSC-1, a cervical cancer cell line robustly expressing SUV420H2. Bar graphs indicate mean ± SD. n = 3 biological replicates each averaged from three technical replicates; differences were assessed by Student’s t test compared with cells transfected with empty vector. **, P < 0.01; ***, P < 0.001. (B) Western blot for H4K20me3 levels in epithelial pancreatic cancer cells with vector control, MT Dead, and wild-type SUV420H2 overexpression. H4 is the loading control. (C) qRT-PCR analysis of expression of epithelial/mesenchymal factors in Capan-1 cells transfected with vector control, MT dead, and wild-type SUV420H2 overexpression plasmid. Data normalized to empty vector transfection level for each gene assayed. Bar graphs indicate mean ± SD. n = 3 biological replicates each averaged from three technical replicates; differences were assessed by Student’s t test compared with cells transfected with empty vector. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (D) Western blots for E-CAD, VIM, and FOXA1 in epithelial pancreatic cancer cell lines exposed to MT dead and wild-type SUV420H2 overexpression plasmid with SUV420H2 knockdown. The loading control is α-TUBULIN. ns, not significant (P ≥ 0.05).