Cst 2965

Linsitinib and JSH‐23 were purchased from Selleckchem Co. (Houston, TX). Stock solutions with a concentration of 10 mM were prepared and stored at −20°C.
Antibodies against phospho‐IGF1R (CST‐3918), total‐IGF1R (CST‐9750), phospho‐p65 (CST‐3033), total‐p65 (CST‐8242), phospho‐Akt308 (CST‐2965), phospho‐Akt473 (CST‐9271), total‐Akt (CST‐1085), phospho‐mTOR (CST‐5536), total‐mTOR (CST‐2983), phospho‐ERK1/2 (CST‐9102), total‐ERK1/2 (CST‐4376), and PARP (CST‐1078) were purchased from Cell Signaling Technology (Danvers, MA, USA). Cleaved Caspase‐3 (25546‐1‐AP) were purchased from Protein Technology (Tucson, AZ) and Tubulin (ARH4207) were purchased from AR (San Diego, CA). HRP‐conjugated goat anti‐mouse and goat anti‐rabbit antibodies were from Santa Cruz Biotechnology (Dallas, TX). MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide] was obtained from Sigma‐Aldrich.

Freshly harvested tissues were fixed in 4% paraformaldehyde (PFA) overnight, and IHC was performed using a SignalStain Citrate unmasking solution (Cell Signaling, 14746) and SignalStain Boost detection reagent (Cell Signaling, 8114) according to the manufacturer’s instructions. Primary antibodies used were rabbit anti-Fgf2 (SC 365106, 1:200; Santa Cruz Biotechnology), rabbit anti-pAkt³⁰⁸ (CST2965, 1:250; Cell Signaling), and rabbit anti–p-Erk (CST9101, 1:100; Cell Signaling).