Dna methyltransferase 1 dnmt1

Western blot (WB) was carried out as previously described [19 (link)]. SDS-PAGE gels (12%) were used to isolate cell lysates, which were subsequently blotted on nitrocellulose membranes and probed with primary antibodies. Primary antibodies against IDH1 (Cell Signaling Technology, Danvers, MA, USA), DNA-methyltransferase 1 (DNMT1; Abcam, Shanghai, China), IDH1 R132H (Dianova, Hamburg, Germany), TET2, Fibulin-5 (Abcam) and actin (Santa Cruz Technology, Dallas, TX, USA) were used in this study. Procedures were carried out independently in triplicate.

The generation of p18^-/-, p18^+/-, p18^-/-;Brca1^MGKO (p18^-/-;Brca1^f/f;MMTV-Cre or p18^-/-;Brca1^f/-;MMTV-Cre), p18^-/-;Brca1^+/-, p18^+/-;Brca1^+/-, GATA3^f/f, GATA3^+/-, p18^+/-;GATA3^+/-, and p18^-/-;GATA3^+/- mice has been previously described 7 (link), 15 (link), 20 (link), 48 (link). The Institutional Animal Care and Use Committee at the University of Miami and Shenzhen University approved all animal procedures. Histopathology and immunohistochemistry (IHC) were performed as previously described 7 (link), 15 (link), 20 (link). The primary antibodies used were: BRCA1, E-cadherin (E-Cad), Vimentin (Vim), FRA1 (Cell Signaling), Fibronectin (Fn), TWIST, DNA Methyltransferase 1 (DNMT1) (Abcam), CD29, GATA3 (Santa Cruz). Immunocomplexes were detected using the Vectastain ABC DAB kit according to the manufacturer's instructions (Vector Laboratories). The positive results of IHC were quantified by H-score, as previously described 49 (link), 50 (link).