Anti s6 p ser235 236

Protein analysis was performed as previously described (Chen et al., 2021a (link)). In brief, total cell lysates were prepared in lysis buffer containing 8 M urea, 2 M thiourea, 3% SDS, 75 mM DTT, 0.05 M Tris-HCl [pH 6.8], and 0.03% bromophenol blue. Lysates from equal numbers of cells for each condition were subjected to SDS-PAGE and analyzed by Western blotting using the following antibodies: anti-IP₃R1 (Ouyang et al., 2014 (link)), anti-IP₃R2 (Ouyang et al., 2014 (link)), anti-IP₃R3 (BD Biosciences, 610312), anti-MEF2C (Abcam, ab211493), anti-c-Myc (Cell Signaling Technologies, 5605), anti-panAkt (Cell Signaling Technologies, 4691), anti-Akt p-Ser473 (Cell Signaling Technologies, 4060), anti-S6 p-Ser235/236 (Cell Signaling Technologies, 4858), anti-β-actin (Santa cruz, sc-47778), and Total Oxphos Rodent WB antibody cocktail (Abcam, ab110413). Proteins were visualized using an HPR-conjugated anti-mouse or anti-rabbit secondary antibody (Cell Signaling Technologies) and chemiluminescent ECL reagent (Thermo Fisher). Densitometric quantification was performed using ImageJ.