Anti c myc

Protein samples were extracted from mammospheres and cancer cells using lysis buffer. After electrophoresis using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), the proteins were transferred to a PVDF membrane (Millipore, Burlington, MA, USA). The membrane was blocked with an Odyssey blocking buffer for 1 h at room temperature and then incubated overnight with primary antibodies. The antibodies were anti-c-Myc (551101, BD, San Jose, CA, USA); anti-Oct4 (LF-MA30482) and anti-GAPDH (LF-PA0018) (AbFrontier, Seoul, Korea); anti-YAP (sc-101199), anti-Lamin B (sc-365962), anti-CD44 (sc-7297), and anti-Sox2 (sc-365923) (Santa Cruz Biotechnology, Dallas, TX, USA); and anti-GR (#12041, Cell Signaling Technology, Danvers, MA, USA). After membranes were washed three times using Tris-buffered saline/Tween 20, all membranes were incubated with IRDye 680RD- and IRDye 800W-labeled secondary antibodies for 1 h at room temperature, and the signal images were determined with an Odyssey CLx (Li-Cor, Lincoln, NE, USA). Densitometric analysis of western blot has been done using Image Studio Ver 5.2 program of Odyssey CLx (Li-Cor, Lincoln, NE, USA).