Anti bach1 antibody

Normal and wound mouse skin samples were paraffin-embedded after being fixed in the 10% formalin solution. To observe wound re-epithelialization and collagen deposition, tissue sections were stained with hematoxylin and eosin (HE), and Masson trichrome respectively. Angiogenesis was assessed by using CD31 immunohistochemical (IHC) staining. Primary antibodies were applied to the sections before being incubated with peroxidase-conjugated secondary antibodies (Cell Signaling, MA). The staining color was visualized using the DAB Peroxidase Substrate Kit (Maxin, China) and was photographed using a SOPTOP CX40 microscope (Shanghai, China)
The following primary antibodies were utilized: anti-CD31 antibody, anti-GPX4 antibody, anti-NFE2L2 antibody, anti-PTGS2 antibody, anti-p-AKT antibody, and anti-BACH1 antibody (Proteintech, USA).

Cell samples and tissue sections were fixed in 4% paraformaldehyde. The samples were then sealed with a 2% (W/V) bovine serum albumin (BSA) solution before being incubated with the primary antibody at 4 °C overnight and the secondary antibody the next day at room temperature. DAPI nuclear dye was utilized to restain the samples. The following primary antibodies were utilized: anti-VEGF antibody, anti-TGF-β antibody, anti-p-AKT antibody, anti-BACH1 antibody (Proteintech, USA), anti-PDGF-B antibody, anti-GPX4 antibody (Abclonal, China).

Cells were divided into five groups: control, BMSC CM, δ-TT-BMSC CM, RSL3, and RSL3 + δ-TT-BMSC CM groups. Cells were gathered and suspended in cell lysis buffer (Cell Signaling Technology, USA) after 24 h treatment, and the supernatant was harvested by centrifugation. Samples were electroblotted onto polyvinylidene fluoride (PVDF) membranes (Bio-Rad, USA) after being separated on a 10% polyacrylamide-SDS gel. PVDF membranes were then blotted with the primary antibody at 4 °C overnight after being blocked in 5% skimmed milk in tris-buffered saline with Tween 20 (TBST) for 1 h. After being washed, PVDF membranes were incubated for 3 h at room temperature with HRP-coupled secondary antibodies (Proteintech, USA) at 1:5000. The ECL WB kit (Proteintech, USA) was used to detect protein signals, which were quantified by ImageLab software. The primary antibodies used in this study were described as follows: anti-VEGF antibody, anti-GAPDH antibody, anti-p-AKT antibody, anti-AKT antibody, anti-PTGS2 antibody, anti-NFE2L2 antibody, anti-BACH1 antibody (Proteintech, USA), anti-PIK3CA antibody, anti-p-PI3K antibody, anti-GPX4 antibody (Abclonal, China).