Anti pstat3

Saw palmetto extract was purchased from Yongyuan Bio-technology, Co., Ltd. (Xi'an, China). TUNEL kit, rabbit anti-B-cell lymphoma-2 (Bcl-2), anti-CD34, anti-MMP-2, anti-PARP, and anti-pSTAT3 antibodies were purchased from the Beyotime Institute of Biotechnology (Shanghai, China).

The protein concentrations were determined using a BCA protein assay kit (Beyotime, Beijing, China). Proteins were electrophoresed on 10% SDS‐polyacrylamide electrophoresis gels and transferred onto a PVDF membrane (Millipore, Merck, Germany). The membranes were blocked in 5% skim milk for 4 h at room temperature. The following were used as primary antibodies: anti‐TLR4 (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐p‐NF‐κB p65, anti‐p‐PI3K, anti‐STAT3, anti‐p‐STAT3 (Beyotime, Beijing, China), anti‐NF‐κB p65, anti‐p‐IκBα, anti‐IκBα, anti‐p‐mTOR, anti‐mTOR, anti‐p‐AKT, anti‐AKT (Cell Signaling Technology, Danvers, MA, USA), anti‐PI3K, anti‐cleaved Caspase 3, anti‐BAX, and anti‐BCL2 (Proteintech, Wuhan, China). Glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH; Proteintech, Wuhan, China) served as the loading control for total proteins. The blots were incubated with the primary antibodies overnight at 4 °C. After washing three times with TBST, the blots were incubated with anti‐mouse secondary antibodies and anti‐rabbit secondary antibodies (Beyotime, Beijing, China) for 1 h at room temperature. Western blots were performed for three independent experiments of each treatment. The intensity of each band was quantified with Image J software (Version 1.52p, NIH).