Ab133644

To assess the protein levels, cell lysates were prepared using lysis buffer containing 20 mm Tris/HCl (pH 7.4), 1% NP‐40, 5 mm EDTA, 100 mm NaF, 2 mm Na₃VO₄, 10 mm Na₄P₂O₇, and protease inhibitor cocktail (#862209; ThermoFisher Scientific, Waltham, MA, USA). Whole‐cell lysates (10 μg) were subjected to SDS/PAGE and immunoblotted with specific antibodies. For immunoblotting, anti‐PPARγ (sc‐7273; Santa Cruz Biotech, Santa Cruz, CA, USA), anti‐C/EBPβ (sc‐7962; Santa Cruz Biotech), C/EBPα (sc‐61; Santa Cruz Biotech), anti‐β‐catenin (#610154; BD Transduction Laboratories, San Jose, CA, USA), anti‐γ‐tubulin (T6557; Sigma‐Aldrich), anti‐FLAG (F1804; Sigma‐Aldrich), phospho‐Y654 FGFR1 (ab59194; Abcam, Cambridge, MA, USA), FGFR1 (#9740; Cell Signaling Technology, Beverly, MA, USA), phospho Y653/Y654 FGFR1‐4 (AF3285; R&D Systems, Minneapolis, MN, USA), FGFR2 (ab109372; Abcam), FGFR3 (ab133644; Abcam), and FGFR4 (ab119378; Abcam) antibodies were used. The proteins were visualized using Clarity™ Western ECL Blotting Substrates (Bio‐Rad, Hercules, CA, USA) or ECL‐Prime (ThermoFisher Scientific) and ChemiDoc Imaging System (Bio‐Rad).

Protein lysates were prepared using RIPA lysis buffer (Beyotime), and quantified using BCA Protein Assay Kit (Beyotime). Proteins were separated by SDS-PAGE and transferred onto nitrocellulose membranes. Blots were then blocked with 5% non-fat milk and incubated with primary antibodies at 4 °C overnight, followed by the incubation with secondary antibody (31430 or 31460, Invitrogen). Signals were detected using ECL substrate (Beyotime). Primary antibodies used in western blot as follows: anti-GPX4 (1:1000, ab125066, Abcam), anti-SLC7A11 (1:1000, ab175186, Abcam), anti-α-SMA (1:2000, ab124964, Abcam), anti-vimentin (1:1000, ab92547, Abcam), anti-FAP (1:2000, PA5-99313, Invitrogen), anti-FGF5 (1:1000, PA5-80630, Invitrogen), anti-Keap1 (1:3000, ab119403, Abcam), anti-Nrf2 (1:2000, PA5-27882, Invitrogen), anti-HO-1 (1:3000, ab68477, Abcam), anti-FGFR1 (1:500, ab76464, Abcam), anti-FGFR2 (1:1000, ab10648, Abcam), anti-FGFR3 (1:1000, ab133644, Abcam), anti-FGFR4 (1:1000, ab178396, Abcam) and anti-β-actin (1:2000, ab8226, Abcam) antibodies. Uncropped western blots were shown in Supplemental materials.

A standard protocol of western blotting was performed. Briefly, cells were lysed using RIPA buffer, and total protein was extracted. After the concentration was determined, the protein was loaded with buffer onto SDS-PAGE gels with subsequent electrophoresis. The protein was then transferred to a PVDF membrane, which was blocked with 5% nonfat milk. Primary antibodies against BCL2L2 (BCL-w) (ab190952; 1:200; Abcam), BCL2L1 (BCL-xL) (ab270253; 1:200; Abcam), FGFRs (ab76464 for FGFR1 at 1:500; ab109372 for FGFR2 at 1:500; ab133644 for FGFR3 at 1:500; ab178396 for FGFR4 at 1:500; Abcam) and NCAM1 (ab9272; 1:500; Abcam) were applied overnight. The corresponding secondary antibodies and ECL were routinely applied. Densitometry was analyzed using ImageStudio software.