Anti mtf 1

HPAEpiCs were lysed in RIPA lysis buffer (ATTO, Tokyo, Japan). The protein extracts were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto a polyvinylidene fluoride membrane (ATTO). After non-specific binding to the membrane was blocked by incubation in 5% skim milk for 1 h, the membrane was incubated overnight with anti-MTF1 (1:1000; Abcam, Cambridge, UK), anti-EGR1 (1:1000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-nuclear receptor superfamily 3, group C, member 1 (NR3C1; 1:1000; Santa Cruz), anti-Lamin B1 (1:1000; Abcam), and anti-α-tubulin (1:1000; Santa Cruz) antibodies. An appropriate horseradish peroxidase-conjugated secondary antibody (1:2000; Cell Signaling Technology, Danvers, MA, USA) was used to bind to the primary antibodies, and the protein bands were imaged using the ChemiDoc Touch Imaging System (Bio-Rad Laboratories).