Figure 6.
Geminin Favors the Formation of B1 Cell-Containing Clones at E14.5
(A) Experimental schematic. The Geminin and Cre plasmids were co-electroporated in utero in MADM-11TG/GT at E13.5 and E14.5, and V-SVZ WMs were analyzed at P15–P20.
(B) Airyscan confocal image of a P15 MADM-labeled V-SVZ whole-mount immunostained with EGFP (green), tdTomato (red), and FOP (white) antibodies showing a clone containing 1 GFP+ ependymal cell (arrow) and two B1 cells (one GFP+ and one tdT+, arrowheads). Note that both B1 cells contain a centrosome at the ventricular surface (arrowheads), and they extend a process toward a blood vessel (yellow asterisks). See also Video S3.
(C) Fitting curves of the distribution of clone size according to the number of cells per clone, issued from electroporation of Cre at E13.5 or E14.5 (dotted curves; n = 6 and 16 animals at E13.5 and E14.5, respectively) or co-electroporation of Cre with Geminin at E13.5 or E14.5 (solid curves; n = 8 and 13 animals at E13.5 Geminin and E14.5 Geminin, respectively); p values were determined with a χ2 test for trend; ns, p > 0.05, ∗∗∗p ≤ 0.001.
(D) Mean percentage of all clones generated from co-electroporation of Cre and Geminin at E13.5 and E14.5 and containing either B1 cells only, ependymal cells only, or a mixed population of ependymal cells and B1 cells. Error bars represent the SEM of 73 or 107 clones; the p value was determined with a two-proportion Z test; ns, p > 0.05.
(E) Mean percentage of ependymal and B1 cells in all clones generated from co-electroporation of Cre and Geminin at E13.5 and E14.5. Error bars represent the SEM of 317 or 335 cells, respectively; the p value was determined with a two-proportion Z test; ns, p > 0.05.
(F) Mean percentage of E-E, E-B1, and B1-B1 cell division in clones generated from E14.5 in controls or after overexpression of GemC1 or Geminin. Error bars represent the SEM of 16, 9, and 13 independent animals electroporated with Cre, Cre+GemC1, or Cre+Geminin, respectively; p values were determined with a Mann-Whitney test; ∗∗∗p ≤ 0.001; ∗∗p ≤ 0.01; ∗p ≤ 0.05; ns, p > 0.05.
The scale bar represents 50 μm.