Pirfenidone

WXWH0265 was provided by WuXi AppTec Group (Shanghai, China) and prepared in vivo in 5% dimethyl sulphoxide (DMSO) and 95% double‐distilled water. The DMSO concentration was limited to 0.1% in vitro. The signal transducer and activator of transcription 3 (STAT3) inhibitors, napabucasin (BBI608), fasudil (HY‐10341A), pirfenidone (HY‐B0673) and GSK429286A (HY‐11000), were purchased from MedChemExpress. S7936, Belumosudil (KD025) was purchased from Selleck Chemicals, USA. Clodronate liposomes were obtained from Liposomes (CP‐005‐005). Anti‐STAT3 (# 12640) and anti‐p‐STAT3 (Y705; # 9145) antibodies were purchased from Cell Signaling Technology (Cambridge, MA, USA). Anti‐α‐SMA (# MAB1420) antibodies were obtained from R&D Systems (Minneapolis, MN, USA). Anti‐ROCK1 (Cat# ab134181), anti‐ROCK2 (Cat# ab71598) and anti‐collagen‐Ⅰ (Cat# ab138492) antibodies were purchased from Abcam (Cambridge, UK). Anti‐β‐tubulin antibodies were purchased from Huabio Technology (Beijing, China).

For targeting the stroma, several compounds were added as 2 µM solutions to the OrganoPlate after medium was removed on day 4. The following compounds were used: Halofuginone (MedChem Express, #HY-N1584) as a PSC/CAF and SMAD 2/3 inhibitor; Galunisertib (Selleck Chemicals, #S2230) as TGF-β receptor inhibitor; Vismodegib (MedChem Express, #HY-10440) as a SHH pathway inhibitor, Pirfenidone (MedChem Express, #HY-B0673) as a cell cycle inhibitor of CAFs; and Losartan (MedChem Express, #HY-17512) as a TGF-β ligand inhibitor (16 (link)).