IMAGE

Fig. 5

ID
ZDB-IMAGE-161122-17
Source
Figures for Katz et al., 2016
Image
Figure Caption

Fig. 5

Active Nucleo-Cytoplasmic Shuttling of Ago and miR-9 Impacts the Quiescent State of NSCs

(A) Scheme of the zebrafish TNRC6 protein with the cloned GW-rich Ago binding domain highlighted in purple.

(B) Triple immunofluorescence of GFP (green), Ago (magenta), and PCNA (light blue) with nuclear DAPI counterstaining (dark blue) on a representative cell electroporated with a control construct (her4:gfp, top, yellow arrow) or the dominant-negative TNRC6 (DN-TNRC6) construct (her4:dn-tnrc6a-gfp, bottom, white arrow). Scale bar, 5 μm.

(C) Quantification of the data from (B), showing the percentage of electroporated cells with mostly nuclear (blue) or cytoplasmic (gray) Ago signal; n = 10 brains per condition.

(D) Percentage of cells electroporated with the her4:gfp (top) or her4:dn-tnrc6-gfp construct (bottom) in which miR-9 is detected in the nucleus (dark blue), cytoplasm (gray) or absent (light blue); n = 10 brains per condition.

(E) Triple fluorescent immunostaining of gfap:GFP (green), Ago (magenta), and PCNA (light blue) with nuclear DAPI counterstaining (dark blue) after ventricular injection of LMB (bottom) or a MeOH control (top). Yellow arrowheads highlight NSCs with a strong nuclear Ago signal. Scale bar, 20 μm.

(F and G) Quantification of data from (E) 24 hpi with LMB or MeOH as a control; n = 5 brains per condition. (F) Percentage of NSCs (gfap+) with strong nuclear Ago signal. (G) Percentage of aNSCs (PCNA+, gfap+) among NSCs (gfap+).

(H) Percentage of aNSCs (PCNA+, gfap+) among NSCs (gfap+) upon combination of LMB or MeOH injections with vivoMO injections. n = 4 brains per condition.

(I) Quantification of the data from (B) of the percentage of GFP+ aNSCs out of total GFP+ NSCs; n = 10 brains per condition.

(J) Percentage of aNSCs (MCM2+, GFP+) out of total NSCs (GFP+) after electroporation with the control her4:gfp or her4:dn-tnrc6-gfp construct and subsequent MO injections. n = 7 brains per condition. p < 0.05, ∗∗p < 0.01; one-way ANOVA with Bonferroni post hoc correction. Data are represented as mean ± 95% CI.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Cell Rep.