Figure 3: Identification of S-phase NPC nuclei by PCNA immunostaining.
From: Neural stem and progenitor cells shorten S-phase on commitment to neuron production
(a, b, c, d) Pax6 (magenta) or Tbr2 (magenta), PCNA (blue) and DAPI staining (c, d, white). Individual nuclei indicated by arrowheads in (a) and (b) are shown at higher magnification in (c) and (d), respectively; white arrowheads, S-phase nuclei showing punctate PCNA immunoreactivity; yellow arrowheads, G1- or G2-phase nuclei showing homogeneous PCNA immunoreactivity. All Pax6+ nuclei (a) and all Tbr2+ nuclei (b) that show punctate PCNA immunoreactivity are indicated by white dots in the respective right panel. Scale bars, 50 μm (a, b) and 10 μm (c, d). (e) Percentage of AP and BP nuclei that are in S-phase as revealed by PCNA immunostaining. Data show the percentage value of (BP) total Tbr2+ nuclei in VZ+SVZ showing punctate PCNA immunoreactivity, corrected for the contribution of neurons, and of (AP) total Pax6+ nuclei in VZ+SVZ showing punctate PCNA immunoreactivity, corrected for the contribution of BPs, and are the mean of four 225-μm-wide fields, each from a different brain and litter; error bars indicate s.d.; **P<0.01. (a–e) Images in (a–d) and data in (e) are from embryos subjected to cumulative EdU labelling for 5 and 3–9 h, respectively.
