Western blot assay was performed as previously described. 24 Briefly, the total protein of glioma tissues was extracted using a protein extraction kit (Beyotime). Equivalent amounts of protein (25‐50 μg) were then electrophoresed and transferred to polyvinylidene fluoride membranes (0.45 μm; Millipore). After being blocked, membranes were incubated with antibodies against ATF6 (24169‐1‐AP, Proteintech), EIF2α (sc‐133132, Santa), p‐EIF2α (# 3398S, CST), p‐IRE1α (sc‐390960, Santa) and GAPDH (60004‐1‐Ig, Proteintech) at 4°C for 16 hours. The membranes were then incubated with appropriate secondary antibodies (ProteinTech). Protein bands of interest were detected and quantified using a chemiluminescence kit (Beyotime), the ChemiDoc™ Touch detection system (Bio‐Rad Laboratories) and Image J software (National Institutes of Health).

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