Anti cct2

Co-immunoprecipitation (co-IP) was performed as described previously^{20 (link),26 (link),49 (link)}. The reagents used included protein A/G-Sepharose (Life sciences, Oslo, Norway) and Western/IP lysis buffer (Beyotime, Haimen, China). The antibodies used for detecting the interactions for endogenous and exogenous proteins were anti-YAP (CST, #12395), anti-TFCP2 (Abcam, #ab180033), anti-PCBP2 (Abcam, #ab184962), anti-CCT3 (Abcam, #ab174255), anti-βTrCP (CST, #4394), anti-IgG (CST,#5415,or CST, #3900), anti-AGPAT1(Abcam, #ab235328), anti-LAMTOR1 (CST, #8975), anti-CCT6A (Abcam, #ab110905), anti-CCT2 (Abcam,#ab92746), anti-FLAG (CST, #8146), anti-HA (CST, #2367), or anti-MYC (CST, #2276).

Cell lysis, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), total protein staining, gel visualization, and gel band quantification were performed as previously described (43 (link)). Lysates were collected from cells grown in 2D culture flasks or 24-well ULA plates. Total protein concentration was determined using the Pierce BCA Protein Assay Kit (Thermo Scientific) following the manufacturer’s protocol. anti-CCT2 (ab109184) and anti-FLAG (ab1162) antibodies were obtained from Abcam, anti-CCT3 (MA5-27872) antibodies were from Invitrogen, and anti-CCT-beta (MAB10050) antibodies were from Millipore. Note that anti-CCT2 (Abcam) and anti-CCT-beta (Millipore) antibodies target the N-terminal amino acids 1-100 and C-terminal amino acids of human CCT-beta, respectively. Secondary antibodies used were IRDye 800CW and IRDye 680CW (LI-COR).