Fig. 4. SHH induces proliferation of nestin-positive cells in the postnatal mouse cerebral neocortex. (A,B) Co-labeling of P3 mouse cortical dissociated cells with anti-nestin and anti-BrdU antibodies. In both control (A) and SHH-treated (B) samples, dividing cells express cytoplasmic nestin (arrows). Nuclei are counterstained with DAPI. Response to SHH induction in dissociated cells included upregulation of Ptch1 (not shown). (C,D) Labeling of dissociated cells with anti-GFAP and anti-BrdU antibodies. BrdU-labeled cells (arrows) do not express GFAP in control (C) or SHH-treated (D) samples. Nuclei are counterstained with DAPI. (E) Quantification of labeling results with dissociated cells. Numbers are percentages of single- or double-labeled cells±s.e.m. over the total number of cells as seen by DAPI staining. n>1000 cells counted for each sample.