Fig. 5

Exosomes containing p62-labeled autophagosomes aggravate inflammation and further stimulate the RIP1/RIP3 pathway to block autophagosome degradation after CIRI. a Immunofluorescence images showing the co-location of p62-labeld autophagosomes and CD63-labeled exosomes in the cerebral cortex after CIRI and Mdivi-1 treatment (bar = 20 μm, n = 5/group). b p62 expression in exosomes after CIRI or OGD/R and Mdivi-1 treatment or Drp1 shRNA (n = 5/group). c p62 expression in exosomes after Drp1 activation (S616A) and Nec1 treatment (10 μmol/L and 50 μmol/L, n = 5/group). d TNF-α and IL-1β levels in the culture supernatant of SH-SY5Y cells after being stimulated with normal or CIRI-derived exosomes (n = 5/group). e Western blotting analysis and Co-IP assay showing RIP3 phosphorylation and RIP1-RIP3 binding ability in SH-SY5Y cells after OGD/R and CIRI-derived exosomes stimulation (n = 5/group). f Immunofluorescence images showing autophagy flux labeled by mRFP-GFP-LC3 in SH-SY5Y cells after OGD/R, CIRI-derived exosomes stimulation, and Nec1 treatment (10 μmol/L and 50 μmol/L, bar = 10 μm, n = 5/group). ^*P < 0.05, compared with normal group; ^#P < 0.05, compared with CIRI or OGD/R group; ^&P < 0.05, compared with CIRI-Exo. group; ^@P < 0.05, compared with CIRI-Exo. + Nec1 (10 μmol/L) group. CIRI cerebral ischemia–reperfusion injury, Co-IP co-immunoprecipitation, Exo. exosomes, OGD/R oxygen–glucose deprivation/reoxygenation treatment