Figure 2. NeuN⁺ Cells Are the Short-Lived, Differentiated Progeny of Cycling DCX⁺ Cells.

(A) Experimental design for (B–D). Ptc mice were injected with 30 mg/kg EdU and sacrificed at successive time points thereafter.

(B) The frequency of all EdU⁺ cells as well as DCX⁺, NeuN⁺, and Sox2⁺ cells that were also EdU⁺ was quantified in primary tumor sections at each postinjection time point (n = 3 per group, mean ± SEM).

(C) Representative immunofluorescence images of DCX and EdU at 3 hr and 14 days postinjection. The arrowhead indicates a rare EdU⁺ label-retaining cell. Scale bars, 20 μm.

(D) Representative immunofluorescence images of NeuN and EdU at 3 hr and 3 days postinjection. Scale bars, 20 μm.

(E) Representative image of activated caspase 3⁺ (AC3) NeuN⁺ cells (arrowhead). The frequency of AC3 events in all cells, NeuN⁺ cells, and Sox2⁺ primary tumor cells is quantified (n = 3 [all], n = 3 [Sox2], n = 8 [NeuN], mean ± SEM, two-tailed unpaired t test [NeuN versus all and NeuN versus Sox2]). DAPI is shown in white. Scale bars, 14 μm.

(F) Representative image of TUNEL staining in NeuN⁺ cells (arrowhead). The frequency of TUNEL events in all, NeuN⁺ cells, and Sox2⁺ primary tumor cells is quantified (n = 3 per group, mean ± SEM, two-tailed unpaired t test [NeuN versus all and NeuN versus Sox]). DAPI is shown in white. Scale bars, 14 μm.

See also Figure S2.