Anti fbp1 antibody

SKOV-3 cells were harvested and lysed in lysis buffer (150 mM NaCl, 50 mM Tris-HCl, 1 mM EDTA [pH 7.5] and 1% NP-40) containing protease inhibitor (Roche, CA, U.S.A.). Whole cell lysates were incubated with an anti-FBP1 antibody, anti-EZH2 antibody or isotype IgG (Abcam, Cambridge, U.K.) at 4°C for 2 h followed by incubation with prepared Protein A+G agarose beads (Santa Cruz, CA, U.S.A.) at 4°C overnight with rotation. After washing with lysis buffer, the precipitates were eluted in SDS-PAGE loading buffer by boiling for 5 min. The supernatants were then resolved by SDS-PAGE and transferred to PVDF membranes. Immunoblotting using appropriate antibodies was conducted using a standard Western blot protocol [47 (link)].

After fixation in 4% formalin and subsequent embedding in paraffin, lung tissues were sectioned at a 3‐μm thickness and stained with anti‐Fbp1 antibody (1:200, Abcam). ImageJ software (National Institutes of Health) was used to assess protein expression.