Figure 2: Cumulative EdU labelling of proliferative and neurogenic APs and BPs.
From: Neural stem and progenitor cells shorten S-phase on commitment to neuron production
(a, b) Pax6 (magenta), Tbr2 (blue), Tis21-GFP (green) and EdU (white) staining after cumulative EdU labelling for 9 h. Individual AP nuclei indicated by arrows/arrowheads in (a) are shown at higher magnification in (b) (asterisks); white arrowheads, Pax6+/Tbr2−/Tis21-GFP−/EdU− nucleus; white arrows, Pax6+/Tbr2−/Tis21-GFP−/EdU+ nucleus; yellow arrowheads, Pax6+/Tbr2−/Tis21-GFP+/EdU− nucleus; yellow arrows, Pax6+/Tbr2−/Tis21-GFP+/EdU+ nucleus. VZ and SVZ are indicated on the left. Scale bars, 50 μm (a) and 10 μm (b). (c, d) Tbr2 (magenta), Tbr1 (blue), Tis21-GFP (green) and EdU (white) staining after cumulative EdU labelling for 9 h. Individual BP nuclei indicated by arrows/arrowheads in (c) are shown at higher magnification in (d) (asterisks); white arrowheads, Tbr2+/Tbr1−/Tis21-GFP−/EdU− nucleus; white arrows, Tbr2+/Tbr1−/Tis21-GFP−/EdU+ nucleus; yellow arrowheads, Tbr2+/Tbr1−/Tis21-GFP+/EdU− nucleus; yellow arrows, Tbr2+/Tbr1−/Tis21-GFP+/EdU+ nucleus. VZ and SVZ are indicated on the left. Scale bars, 50 μm (c) and 10 μm (d). (e, f) Proportion of EdU-labelled NPC nuclei (EdU labelling index) after cumulative EdU labelling for 0.5, 1, 2, 3, 5, 9, 12, 18, 24 and 29 h. The EdU labelling index was separately determined for the various AP (Pax6/Tbr2/GFP/EdU staining as in (a)) and BP (Tbr2/Tbr1/GFP/EdU staining as in (c)) populations, as follows: total APs (e, Tbr2−/Pax6+, blue circles), total BPs (e, Tbr1−/Tbr2+, red squares), proliferative APs (f, Tbr2−/Pax6+/Tis21-GFP−, blue diamonds), BP-genic/neurogenic APs (f, Tbr2−/Pax6+/Tis21-GFP+, purple squares), proliferative BPs (f, Tbr1−/Tbr2+/Tis21-GFP−, red triangles) and neurogenic BPs (f, Tbr1−/Tbr2+/Tis21-GFP+, green crosses). Data are the mean of three 225-μm-wide fields (except for the 1-, 9- and 29-h time points, which are from five, two and one field(s), respectively), each from a different brain and litter; error bars indicate s.e.m. or the range of the two individual values. Dashed lines and colour-coded arrowheads indicate the growth fraction; colour-coded arrows indicate the time point at which the labelling index reaches a plateau (TC–TS).
