Anti hsp90

The following antibodies were used in the study: anti‐SFRP2 (Santa Cruz, sc‐365,524, 1:300), anti‐THBS2 (Abcam, ab84469, 1:500), anti‐HSP90 (ABclonal, A5027, 1:2000) and anti‐GAPDH (ZSGB‐BIO, TA‐08, 1:2000). Secondary antibodies (Jackson Immuno Research Laboratories, West Grove, PA, USA) were diluted 1:4000 using 5% skim milk in Tris‐buffered saline.

Immunoblotting analysis was performed using the following antibodies: anti-Phospho-p70 S6 Kinase (Thr398) (cat# 9324), anti-p70 S6 Kinase (cat# 9202), anti-p44/42 (Erk1/2) (cat# 9102), anti-ATF-4 (cat# 11815), anti-p38 (cat# 8690) from CST, anti-Flag (Sigma-Aldrich, cat#11815), anti-Xdh (Proteintech, cat# 55156-1-AP), anti-HSP90 (Abclonal, cat# A5027). Briefly, protein concentration in samples was quantified by Quick Start™ Bradford 1× Dye Reagent (BIO-RAD, cat# 5000205); then the samples were equally loaded for electrophoresis and membrane transfer. The membrane was blocked with 5% (m/v) milk in TBST (0.1% Tween 20) RT for 30 min before being incubated with primary antibody overnight at 4 °C. The membrane was washed three times with TBST and incubated with the corresponding secondary antibody RT for 1 h. The membrane was washed three times before exposure using Immobilon Western Chemilum HRP Substrate (Millipore Sigma).