Rabbit anti foxa1 antibody

Immunofluorescence analysis was performed as described previously in Material and Methods. The cell slides were sterilized by immersion in 70% alcohol for 30 min. HEK293T and Hela cells were seeded in a 24-well plate with a cell slide at a density of 1×10⁴ cells per well. The cell slides were removed after 24 h of transfection. Finally, the expression and localization of Foxa1 in cells were examined by immunofluorescent staining. Cell slides were treated in PBS containing 5% bovine serum albumin (BSA) and 0.5% Triton X-100 for 30 min to inhibit non-specific binding, followed by a 2-h incubation with rabbit anti-Foxa1 antibody (Abcam, 1:400 dilution) at room temperature. After three PBS washes, the slides were incubated with Alexa Fluor546-labeled anti-rabbit IgG (H+L) secondary antibody (Abcam, 1:500 dilution) in the dark for 2 h. The results were obtained using a Leica DMI8 fluorescence microscope (Leica).

Rat footpads were dissected in cold (4°C) 1× phosphate buffered saline (PBS) and fixed in 4% paraformaldehyde solution for 24 h at room temperature (24°C). Paraffin-embedded tissues were sectioned, deparaffinized, and hydrated. The HE staining kit (Beyotime, China) was used for the staining. Briefly, the sections (5-μm-thick) were stained with hematoxylin for 2 min and then immersed in the acidic liquid alcohol for 30 s. The images were obtained using an inverted microscope after staining with eosin for 2 min and dehydrated with ethanol (95, 100%). The samples were blocked with QuickBlock™ Blocking Buffer for Immunostaining (Beyotime) for 10 min, labelled with rabbit anti-Foxa1 antibody (Abcam, 1:400 dilution, UK), mouse monoclonal anti-NKA α2 (1:200 dilution, Santa Cruz, USA) for 8 h at 4°C, and then incubated with Alexa Fluor 546-labeled anti-rabbit IgG (H+L) secondary antibodies (1:500 dilution, Abcam) for 2 h in the dark. For counterstaining, the sections were incubated with 4′,6-diamidino-2-phenylindole (DAPI, Beyotime) in the dark for 1 min at room temperature. Finally, the sections were sealed with an anti-fluorescence quencher. The results were obtained using a Leica DMI8 fluorescence microscope (Leica, Germany).