Ab106836

MH7A cells were seeded into 12‐well slides and cultured in a DMEM medium containing 10% FBS. When the cells reached 80% confluence, they were treated with TNF‐α or PBS (Ctrl) combined or not combined with TG for 1 h. Next, the treated cells were fixed with 4% paraformaldehyde, blocked with PBS containing 5% goat serum, and permeabilized with 0.2% Triton X‐100/PBS for 30 min. The cells were then hybridized with a cy3‐labled Lnc‐ENST00000602558 probe (GenePharma) overnight. The next morning, the cells were washed with 20×SSC eluant (Servicebio) and then incubated with IGF1 antibody (Abcam, ab106836, 1:500) overnight at 4°C. The next morning, the slides were re‐warmed for 1 h at room temperature, and then incubated with a secondary conjugated fluorescent dye (Abcam, ab150129, 1:1000) at 37° for 1 h. After counterstaining with DAPI (Abcam, ab228549, 1:5000), images were captured with a Nikon Eclipse TS100 microscope equipped with Micro‐Manager 1.4.22 acquisition software.

RIPA lysis (Pierce) was employed for extracting proteins. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred on PVDF membranes. Following Bloced by skim milk, the membranes were immunoblotted with primary antibodies including IGF1 (1.5 µg/mL, ab106836, Abcam) and β-actin (1 µg/mL, ab8226, Abcam). Post washing by Tris-buffer with Tween 20, secondary antibody goat anti-rabbit IgG- horseradish peroxidase (HRP) conjugate (0.5 µg/mL, ab6721, Abcam) was filled in and incubated for 1 hour. Subsequently, an ECL Western Blotting Substrate Kit (TransGen Biotech, China) was applied to visualising the protein bands.