Sc 222227

The PTP1B inhibitor, sc-222227 (Santa Cruz Biotechnology, Inc.), was dissolved in DMSO and diluted with 0.9 saline to a final concentration of < 0.1% DMSO. Anesthetized rats were placed in a stereotaxic frame, and sc-222227 (5 and 10 μM) was injected into the right cerebral ventricle with a Hamilton syringe using the following coordinates: 3 mm rostral to the bregma; 2 mm lateral to the midline; and 2 mm ventral to the skull surface. The injection rate was 0.2 μL/min, and the needle was left in place for 5 min after injection before being slowly withdrawn. After drug administration, MCAO and reperfusion were performed immediately, as described above. Both sham and IR groups were administered vehicle (DMSO) intracerebroventricularly.

The PTP1B inhibitor, sc-222227, was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The autophagy inhibitor, N(3)-methyladenine (3-MA) and the ER stress inhibitor, 4-phenylbutylamine (4-PBA), were purchased from Sigma (Sigma, St. Louis, MO, USA). The protein kinase R-like endoplasmic reticulum kinase (PERK) activator, CCT020312, was purchased from Merck Millipore (Billerica, MA, USA). The PERK inhibitor, GSK2606414, was purchased from Sigma. Antibodies were obtained from the following sources: CD11b/c, PTP1B, inositol requiring enzyme-1 (IRE1), p-IRE1, activating transcription factor 6 (ATF6), neuronal nuclear antigen (NeuN), ionized calcium binding adapter molecule 1 (Iba-1), heavy chain binding protein (Bip), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-actin were purchased from Abcam (Cambridge, MA, USA); LC3-I/II and beclin-1 were purchased from Cell Signaling (Danvers, MA, USA); glial fibrillary acidic protein (GFAP) was purchased from Santa Cruz Biotechnology, Inc.; primary antibody for p-PERK (Thr982), PERK, p-eIF2α (Ser51), eIF2α, CD16/32, CD206, and secondary antibodies were purchased from Invitrogen (Carlsbad, CA, USA); and donkey anti-rabbit IgG, rabbit anti-goat IgG, and goat anti-mouse IgG were purchased from Invitrogen.