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After blocking in 5% skimmed milk powder for 2 hours at room temperature, the samples were incubated overnight at 4C with primary antibodies, including ERK1/2 (#9102), p-ERK1/2 (#9101), JNK1/2 (#9252), p-JNK (#9251), p38 MAPK (#9212), and p-p38 MAPK (#9211), which were obtained from Cell Signaling Technology (Beverly, MA, USA), as well as VEGF-a (sc-7269) and Tubulin (sc-8035), which were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA)

The SP-9000 SPlink Detection Kit (Biotin-Streptavidin HRP Detection Systems, ZSGB-BIO, Beijing, China) was used to detect antibody binding

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Briefly, recombinant PDI protein (0.4 M) was incubated with indicated compounds at 37 C for 1 h in sodium phosphate buffer (100 mM sodium phosphate, 2 mM EDTA and 8 M DTT, pH 7.0)