Fig 2: Variation in CRT protein level and mRNA expression of CRT, caspase-8, and caspase-3 in the cortexes of MCAO mice. The ipsilateral cortexes of the sham and MCAO mice were collected at 3, 6, 12, and 24 h after reperfusion. (a) CRT mRNA expression detected by real-time PCR and (b1, 2) CRT protein levels detected by Western blotting. (c) Caspase-8 and (d) caspase-3 mRNA detected by real-time PCR. * P < 0.05, ** P < 0.01, and *** P < 0.0001 versus sham. n = 6 repeats.

Fig 3: CRT binds to FasL on the surface of neurons after ischemia. (a) Immunofluorescent staining of the cortexes of sham and MCAO mice. (b) Confocal microscopy images of immunostained normal SH-SY5Y cells and OGD-exposed SH-SY5Y cells at 3 h after reperfusion. (c1, 2) Co-IP of CRT and FasL in the cortexes of sham and MCAO mice. (d1, 2) Co-IP of Fas and FasL in normal SH-SY5Y cells and OGD-exposed SH-SY5Y cells cultured with or without 40 ng/mL CRT. ** P < 0.01, *** P < 0.0001 versus control, and ### P < 0.0001 versus OGD group. n = 6 repeats.

Fig 4: CRT prevents apoptosis among OGD-exposed SH-SY5Y cells and downregulates the activity of caspase-8 and caspase-3. SH-SY5Y cells were cultured in medium with or without 40 ng/mL CRT after OGD. (a1, 2) Apoptosis rate of OGD-exposed SH-SY5Y cells as detected by flow cytometry at 6 h after reperfusion and activity of (b) caspase-8 and (c) caspase-3 as detected using caspase activity assay kits at 3, 6, 12, and 24 h after reperfusion. Control: normal SH-SY5Y cells. ** P < 0.01, *** P < 0.0001 versus control and # P < 0.05 versus OGD group. n = 6 repeats.