Anti hnf4α antibody

Pancreatic tissue sections from βUCP2Tg mice and in vitro-cultured human islets were immunostained with anti-insulin (Abcam), anti-UCP2 (Santa Cruz), anti-AldB (Proteintech), or anti-BrdU (Dako, Tokyo, Japan) antibodies. Images were acquired as described elsewhere (Shirakawa et al., 2013 (link)). For immunofluorescence cytochemistry (IFCC), 1 × 10⁴ MIN6-M9 cells were seeded on poly-L-lysine-coated chamber slides (Nunk, #154534PK) and infected with Ad-LacZ or Ad-Ucp2 at an MOI of 500 for 48 h. Cells were fixed with 4% PFA and were permeabilized with 0.01% Triton X-100 in PBS. Anti-HNF4α antibody was purchased from Abcam (#ab201460). Images were acquired using an FV1000-D confocal laser scanning microscope (Olympus, Japan). Super resolution STED microscopic images were acquired using TCS SP8 STED (Leica Microsystems, Germany).

Total cell proteins were obtained with the Western IP lysis buffer (Beyotime Biotechnology). BCA assay, SDS–PAGE, and Western blotting were performed using a standard procedure. The primary antibodies against HNF1α, FOXA2, and ERα were obtained from CST (Boston, MA, USA). The anti-GAPDH antibody was purchased from Beyotime Biotechnology. The anti-HNF4α antibody was obtained from Abcam (Cambridge, UK). The second antibodies HRP-labeled Goat anti-Mouse IgG or HRP-labeled Goat anti-Rabbit IgG were obtained from Beyotime Biotechnology. All antibodies were diluted at the ratio of 1:1000. The protein bands were visualized by ultrasensitive chemiluminescence (ECL; Millipore, MA, USA) with the ChemiDoc XRS⁺ Imager (Bio-Rad, Hercules, CA, USA) and analyzed by Image J software (NIH, Bethesda, MA, USA).