Non targeting control sirna

For siRNA knockdown, GMSCs (0.2 × 10⁶) were seeded on a six-well culture plate. Fas, FAP-1, Cav-1, SNAP25, and VAMP5 siRNAs (Santa Cruz Biotechnology) were used to treat the GMSCs according to the manufacturer’s instructions. Nontargeting control siRNAs (Santa Cruz Biotechnology) were used as negative controls. Fap-1 CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) knockout plasmid (sc-422505, Santa Cruz Biotechnology) was used to knock out Fap-1 expression in GMSCs according to the manufacturer’s instructions. Briefly, GMSCs (0.2 × 10⁶) were seeded on a six-well culture plate. The cells were allowed to grow to 40 to 80% confluence and then transfected with Fap-1 CRISPR/Cas9 knockout plasmids using Lipofectamine LTX with Plus reagent (Life Technologies) according to the manufacturer’s instructions. Scrambled guide RNA CRISPR/Cas9 plasmids were used as a negative control. The efficiency of siRNA knockdown and CRISPR/Cas9 knockout was confirmed by Western blot analysis. For cytokine treatments, GMSCs were treated with different concentrations of IFN-γ and TNF-α (Peprotech; 0, 20, 50, 100, and 200 ng/ml) or TNF-α (20 ng/ml) for 24 hours. After transfection or cytokine treatment, cells were used for protein extraction for Western immunoblotting, and the culture supernatants were used for ELISA.