Fig 1.
Experimental protocol and ischemic injury location.
Experimental protocol in days (A). The vasoconstrictor Endothelin-1 was injected into the striatum to induce ischemia. The ischemic area in the striatum is circled in black (B).
Fig 2.
Effect of ND-13 treatment on functional recovery after focal ischemic injury.
ND-13 treatment significantly improved time spent crossing the bridge in the elevated bridge test, two days after injury, compared to control (A, p<0.05). The time to descend from a vertical pole in the pole test also decreased following ND-13 treatment compared to control (B, p<0.05). The motor asymmetry in the cylinder test decreased 2 days after ischemic injury (C, p<0.05). ND-13 effect was still consistent 21 days after ischemic injury in all behavioral tests, that is, mice treated with ND-13 perform better than control mice. (Data is shown as mean ± SEM).
Fig 3.
DJ-1 KO mice show higher sensitivity and less spontaneous recovery after ischemic injury compared to C57BL/6 mice.
DJ-1 KO mice show slower spontaneous recovery compared to C57BL/6 mice and are more sensitive to ischemic insult. DJ-1 KO mice spent significantly more time crossing the beam in the elevated bridge test (A, p<0.05) than wild type mice. DJ-1 KO mice show only 5% recovery from day 2 to days 7 after injury, compared to C57BL/6 mice that improved by 20% over the same period of time. Improvement was also noted in cylinder test 7 days after injury (B). (Data is shown as mean ± SEM).
Fig 4.
Effect of ND-13 treatment on functional recovery of DJ-1 KO mice after ET-1 induced focal ischemic injury.
DJ-1 KO mice treated with ND-13 show improvement 2 days after ischemic injury in the elevated bridge test as they spend less time crossing the beam than wild type mice (A, p<0.05). This effect was consistent for at least two weeks following injury. Improvement was also noted in cylinder test 7 days after injury (B). (Data is shown as mean ± SEM).
Fig 5.
Changes in mitochondrial protein expression levels following ND-13 treatment.
Ischemic wild type mice treated with ND-13 show significant change in protein expression levels compared to saline treated ischemic wild type mice. SDHAF4 protein levels increased after ND-13 treatment (A, FC = 107.4, p<0.001). Ubiquinone protein levels decreased after ND-13 treatment (B, FC = 3.5, p<0.01). Rhot2 protein levels decreased after ND-13 treatment (C, FC = 2.5, p<0.01). Usp35 protein levels decreased after ND-13 treatment (D, FC = 2.4, p<0.01). (Data is shown as mean ± SEM).
Fig 6.
Changes in potassium channel regulators protein expression levels following ND-13 treatment.
Ischemic WT mice treated with ND-13 show significant change in protein expression levels compared to WT mice treated with saline, 2 days after injury. CRBN protein levels decreased after ND-13 treatment (A, FC = 6.7, p<0.01). Kcnip4 protein levels decreased after ND-13 treatment (B, FC = 3.4, p<0.01). (Data is shown as mean ± SEM).