Fig. S5
Related to Figure 4, 6, 7: Control experiments for morpholino knockdown. (A) Knockdown of PI3K α, β or δ isoforms using morpholinos does not rescue bace2-/- dendritic melanophores to the same extent as PI3K γ morpholinos, as shown in Figure 4E. (B) Knockdown of mTOR using mTOR splicing morpholino leads to depletion of mTOR protein in Western blot. Membrane probed with anti-mTOR antibody and β-actin as a loading control. (C) RT-PCR shows knockdown of mTOR using mTOR splicing morpholino leads to abnormal mRNA splicing. mtor fragment is amplified using primers adjacent to morpholino target sites. actin: input control. (D) Western blot shows co-knockdown of insra and insrb using morpholinos leads to decrease insulin receptor protein. Membrane probed with antibody sc-711 recognizing insulin receptor β chain and β-actin as a loading control. (E) Knockdown of insa (Left) and insb (Right) using splicing morpholinos lead to abnormal mRNA splicing and expression in RT-PCR. Fragments are amplified using primers adjacent to morpholino target sites. actin: input control. Scale bars, 100μM. |
Reprinted from Developmental Cell, 45(5), Zhang, Y.M., Zimmer, M.A., Guardia, T., Callahan, S.J., Mondal, C., Di Martino, J., Takagi, T., Fennell, M., Garippa, R., Campbell, N.R., Bravo-Cordero, J.J., White, R.M., Distant Insulin Signaling Regulates Vertebrate Pigmentation through the Sheddase Bace2, 580-594.e7, Copyright (2018) with permission from Elsevier. Full text @ Dev. Cell