Fig. S3
Isolation and differentiation of zebrafish neurospheres, related to Figure 5. (A) Dorsal view of a whole-mount zebrafish adult brain (~10 months). Black dotted lines indicate the anatomical dissection performed to separate different regions of the brain. (B) Schematic representation of neurophere preparation starting from the brain regions indicated in (A). (C) Bright-field image of neurospheres after 4 days in vitro (Div4). (D) Chart representing the number of neurospheres at Div4 generated from neural cells isolated from telencephalon and tectum relative to the cerebellum area. Data are representative of two independent experiments. (E) Immunostaining of GFAP protein and BrdU incorporated in neurospheres at Div4. White arrow indicates GFAP positive cells. (F) Immunolocalization of the indicated proteins in one neurosphere at 4 days in vitro after differentiation (Div4d). Right panels are high magnifications of the regions highlighted in the left image. Arrowheads show the cells labeled with the indicated antibodies and for the expression of the mCherry under the SOX10 promoter sequence. (G) qRT-PCR quantification of the indicated genes expressed in neurosphere cultures at Div4 and at Div4d. Levels are relative to Div2. Data represent the mean ± SEM (n=3).* p<0.05. Scale bars= 50µm. |
Reprinted from Developmental Cell, 32(5), Ristori, E., Lopez-Ramirez, M.A., Narayanan, A., Hill-Teran, G., Moro, A., Calvo, C.F., Thomas, J.L., Nicoli, S., A Dicer-miR-107 Interaction Regulates Biogenesis of Specific miRNAs Crucial for Neurogenesis, 546-60, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell