Anti β tubulin

The cells were fixed in 4% paraformaldehyde for 10 min at −20°C and then permeabilized 10 min with 0.1% Triton X-100 at room temperature. Following blocking in 10% goat serum for 30 min at room temperature, cells were incubated with the primary antibodies anti−CCT5 (Santa Cruz Biotechnology, USA, #SC374554), and anti−β−tubulin (Sungene Biotech, China, #KM9003T) at 4°C overnight. After that, the cells were washed with TBST and incubated 1 h with 1μLAlexa Fluor^®488 donkey anti-mouse lgG at room temperature (life technologies, USA, #A21202). After washing with TBST, nuclei were stained with DAPI (Solarbio, China) for 5 minutes. Images were acquired by confocal microscopy (Leica Microsystem SP8, Wetzlar, Germany). The fluorescence intensity was quantified using ImageJ software.

Western blotting was performed as previously described [17 (link)]. The primary antibodies used were as follows: anti-PTEN (Abcam Cat# ab32199, RRID:AB_777535; Cell Signaling Technology Cat# 9188, RRID:AB_2253290; Proteintech Cat# 60300-1-Ig, RRID:AB_2881415), anti-NGAL (Abcam Cat# ab63929, RRID:AB_1140965), anti-FN (Sigma-Aldrich Cat# F3648, RRID:AB_476976), anti-Col-I (Boster Biological Technology Cat# BA0325, RRID:AB_2891224), anti-α-SMA (Sigma-Aldrich Cat# A5228, RRID:AB_262054), anti-PCNA (Proteintech Cat# Biotin-60097, RRID:AB_2883063), anti-E-Cadherin (Cell Signaling Technology Cat# 14472, RRID:AB_2728770), anti-Flag (Proteintech Cat# 20543-1-AP, RRID:AB_11232216), anti-CHMP2A (Proteintech Cat# 10477-1-AP, RRID:AB_2079470), anti-SQSTM1/p62 (Cell Signaling Technology Cat# 39749, RRID:AB_2799160), anti-LC3B (Cell Signaling Technology Cat# 83506, RRID:AB_2800018), anti-LAMP1 (Cell Signaling Technology Cat# 15665, RRID:AB_2798750), anti-β-tubulin (Tianjin Sungene Biotech Cat# KM9003, RRID:AB_2744678), and anti-GAPDH (Tianjin Sungene Biotech Cat# KM9002, RRID:AB_2721026). Western blotting was performed at least three times, independently. Quantification was performed by measuring the signal intensity using the software ImageJ (National Institutes of Health).