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. Author manuscript; available in PMC: 2022 Mar 25.
Published in final edited form as: Dev Cell. 2015 Feb 5;32(5):546–560. doi: 10.1016/j.devcel.2014.12.013

Figure 5. Loss of the miR-107-dicer interaction promotes neurogenesis and correlates with an increase in miR-9 biogenesis and activity in zebrafish brain-derived neurospheres.

Figure 5

(A) Schematic representation of the neurosphere assay. (B) qRT-PCR of dicer and miR-107 levels in neurospheres at the stages indicated. Expression levels are relative to Div2. Data are represented as mean ± SEM (n=3). *p<0.05. (C) qRT-PCR of dicer and miR-107 levels in neurospheres at Div4d treated as indicated at Div4. Levels are relative to cells transfected with anti-miR-scramble for anti-miR-107 samples and mimic-scrambled for mimic-miR-107 treatments. Values are presented as mean ± SEM (n=3 to 5). *p<0.05. (D) Left panels are confocal images showing neurospheres at Div4 and Div4d stages treated at Div2 and Div4 respectively with the indicated oligonucleotides. Neurospheres at Div4 are stained with the anti-PCNA antibody and neurospheres at Div4d are stained with an anti-HU antibody. Right charts show the quantification of the total number of PCNA and HU positive cells normalized to the total number of DAPI cells per neurospheres. Values are represented as fold change of PCNA/DAPI or HU/DAPI double positive cells in treated neurospheres compared to the respective controls. Data represent the mean ± SEM (n=4 for the PCNA and n=5 for HU immunodetection). *p<0.05. (E) qRT-PCR of indicated miRNAs expressed in neurospheres at Div4d treated at Div4 with anti-miR-107. Levels are relative to the control (anti-miR-scrambled). Data represent the mean ± SEM (n=4). *p<0.05. Scale bars= 50μm.