Anti pycr1

The HCC cell lines used in this study included Huh7 cells and LM3 cells. Because the PYCR1 expression of these two cells were higher than the other cells (Figure S1). The HCC cell lines were obtained from the China Center for Type Culture Collection (Wuhan, China). These cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS; Gibco) and maintained in a humidified incubator with 5% CO₂ at 37 °C.
The following antibodies were used in immunoblotting and immunofluorescence studies: anti-β-actin (HRP-60008; Proteintech Group, China), anti-PYCR1 (131081; Proteintech Group), anti-E-cadherin (PA5-19479; ThermoFisher Scientific, USA), anti-β-catenin (ab32572; Abcam, USA), anti-N-cadherin (ab76011; Abcam), and anti-vimentin (ab92547; Abcam).
Reagents and materials used for the analysis of cell proliferation, migration, and invasion and for RNA-seq included Cell Counting Kit‑8 (CCK‑8; CK04, Dojindo Molecular Technologies, Inc., Rockville, MD, USA), Transwell plates (8 μm pore size; Corning, Inc.), RNA Nano 6000 assay kit (Agilent Technologies, CA, USA), and NEB Next® Ultra™ RNA library prep kit for Illumina® (#E7530L; New England Biolabs, USA).

For immunofluorescence, cells or slides were fixed (4% paraformaldehyde), permeabilized (0.5% Triton X-100), blocked (5% goat serum), and then stained with anti-PYCR1 (1:200, Proteintech, 13108-1-AP), anti-keratin (1:200, Cell Signaling Technology, 4545), anti–pro–surfactant protein C (anti–pro-SPC; 1:250, Abcam, ab90716), and anti–α-SMA (1:500, Servicebio, GB111364) primary antibodies overnight at 4°C. Afterward, samples were probed with secondary goat anti-rabbit or goat anti-mouse IgG antibody conjugated with Alexa Fluor 555 (Invitrogen, A-21428, A-21422) for 1 hour at room temperature in the dark. Nuclei were stained with DAPI (Yeason, 36308ES20). Images were visualized with a fluorescence microscope (Olympus, model IX73) and analyzed via ImageJ.