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Western blotting was performed as previously described [8 (link)]. First, the total cell protein extraction kit (KeyGen Biotechnology, Nanjing, China) was used to isolate the total protein of GSCs. The nuclear and cytoplasmic protein was isolated using a NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific). Then, the proteins were separated by 4 to 20% SDS-PAGE (Genscript, Nanjing, China), transferred onto polyvinylidene difluoride (PVDF) membranes, and blocked with 2% bovine serum albumin (BSA, KeyGen Biotechnology) and incubated with the primary antibodies at 4 °C overnight, followed with secondary antibodies (ProteinTech, Chicago, Illinois, USA) incubation. The chemiluminescence ECL kit (Beyotime Biotechnology, Beijing, China) visualized the bands. All results were quantified by ImageJ software (National Institutes of Health, Bethesda, MD, USA).

The total protein of EOC tissues and cell lines was isolated via a cell protein extraction kit (KeyGen Biotechnology, Nanjing, PR China) according to the manufacturer’s instructions. The BCA kit (Beyotime Biotechnology, Beijing, PR China) was used to determine the concentration. An equivalent amount of protein from each sample was separated by 4 to 20% SDS-PAGE (Genscript, Nanjing, China), transferred to a nitrocellulose membrane and blocked with 2% bovine serum albumin (KeyGen Biotechnology). The membrane was then incubated with primary antibodies against NPTX2, IL6, p-JAK2, JAK2, p-STAT3, STAT3 and β-actin (Abcam Technology, Cambridge, UK) at 4°C overnight, followed with TBST washing and incubated with secondary antibody. The ECL kit (Beyotime) was used to detect the bands on each membrane, and IMAGE J software (National Institutes of Health, Bethesda, MD, USA) was used for quantification.