1. Z-FA-FMK stabilized both SMN-FL and SMN-Δ7 proteins in Myc-SMN2a– and Myc-SMN2d–inducible lines.
Z-FA-FMK stabilized both SMN-FL and SMN-Δ7 proteins in Myc-SMN2a– and Myc-SMN2d–inducible lines. (A) In Myc-SMN2a– and Myc-SMN2d–inducible lines, addition of doxycycline induced a robust expression of Myc-SMN proteins. Scale bar, 50 μm. (B–E) After the withdrawal of doxycycline, inducible cells were treated with Z-FA-FMK, DMSO (vehicle control), and MG132 (positive control) for 12 h to examine the protein degeneration. (B, D) Western blot showing the protein expression of the Myc-SMN2a (B) and Myc-SMN2d (D) at different time points after doxycycline withdrawal in Myc-SMN2a– and Myc-SMN2d–inducible cells, respectively. (C, E) Quantification from independent experiments showing a significant increase in the expression of both Myc-SMN2a (C) and Myc-SMN2d (E) proteins in the Z-FA-FMK–treated (n = 7 and 5) or MG132-treated (n = 3) group compared with DMSO-treated (n = 7 and 5) group. Data were presented as mean ± SEM. *P < 0.05, **P < 0.01, as compared with the DMSO group by two-tailed t test. (F) Western blot with anti-ubiquitin antibody after co-immunoprecipitation of Myc-SMN showing the ubiquitinated Myc-SMN in the MG132-treated group, rather than in the DMSO- or Z-FA-FMK–treated group.
Yiran Wang et al. LSA 2019;2:e
© 2019 Wang et al.