1. Protective effects of cysteine protease inhibitor Z-FA-FMK and E64d in SMNΔ7 mice.
Protective effects of cysteine protease inhibitor Z-FA-FMK and E64d in SMNΔ7 mice. (A) Western blot and quantification revealed that spinal SMN proteins of SMNΔ7 mice at PND4 significantly increased after Z-FA-FMK treatment. n = 4. (B) Immunostaining and quantification revealed that L3–L5 motor neurons labeled with ChAT antibody in SMNΔ7 mice at PND6 significantly increased after Z-FA-FMK treatment. Scale bar, 100 μm. WT control (n = 6), SMA+DMSO–treated (n = 7), SMA+Z-FA-FMK–treated (n = 5) mice. (C) Survival curves of WT control (n = 18), SMA+DMSO–treated (n = 8), and SMA+Z-FA-FMK–treated (n = 15) mice. (D) Body weight of WT control (n = 18), SMA+DMSO–treated (n = 8), and SMA+Z-FA-FMK–treated (n = 15) mice. (E) Immunostaining showing the expression of Isl1/Tau in long-term SMA cultures (6-wk) after the treatment of E64d (10 μM) or DMSO. (F) Quantification revealed a significant increase in the proportion of Isl1+ spinal motor neurons by E64d; n = 3. (G) SMA mice were treated with E64d or DMSO from day 1. Western blot and quantification revealed that spinal SMN protein level of SMNΔ7 mice at PND4 significantly increased after E64d treatment. n = 3. (H) Immunostaining and quantification revealed that L3–L5 motor neurons labeled with ChAT antibody in SMNΔ7 mice at PND6 significantly increased after E64d treatment. Scale bar, 100 μm. WT control (n = 6), SMA+DMSO–treated (n = 5), SMA+E64d–treated (n = 5) mice. (I) Survival curves of WT control (n = 8), SMA+DMSO–treated (n = 6), and SMA+E64d–treated (n = 8) mice. The statistical significance for the Kaplan-Meier survival analysis was determined by Log-rank test. (J) Body weight of WT control (n = 8), SMA+DMSO-treated (n = 6), and SMA+E64d-treated (n = 8) mice. Data were presented as mean ± SEM. **P < 0.01 as compared with the WT mice. #P < 0.05, ##P < 0.01, ###P < 0.001, as compared with the SMA+DMSO group by two-tailed t test.
Yiran Wang et al. LSA 2019;2:e
© 2019 Wang et al.