TMZ and Ara-C treatment of slice co-cultures.

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TMZ and Ara-C treatment of slice co-cultures. TMZ and Ara-C treatment of slice co-cultures. (A) DNA damage responses of mouse IENS-GFP cells following treatment with various does of the antimitotic agents AraC and TMZ. Immunocytochemistry for gamma-H2AX (gH2AX, yellow) and p53 (red). Nuclear counterstaining was performed with DAPI (blue) (B) Quantitation of the mean intensity of the nuclear H2AX foci (arbitrary units). (C) Schematic of the co-culture of IENS-GFP cells in the brain slices, adding pharmacological inhibitors of proliferation. (D) Immunostaining for GFP (green) and Ki67 (magenta) with DAPI (blue) for nuclear counterstaining. (E) Quantification of the percentage of Ki67+ (proliferative) cells using the proliferative inhibitors in IENS-GFP after 24 h. (F) Immunocytochemistry for GFP (green) and pHH3 (mitotic cells) (yellow) in IENS-GFP after treatments. Percentages of pHH3-positive cells are shown in yellow type. (G) Quantification of the percentage of active caspase 3 in the IENS-GFP cells after treatment with Ara-C and TMZ. (H) Immunostaining for active caspase 3 in the IENS-GFP cells in the brain slices after treatments. Scale bars: 20 µm in A and F; 30 µm in D and H. n=3, Student’s t-test, *P<0.05, **P<0.01. Maria Angeles Marques-Torrejon et al. Dis. Model. Mech. 2018;11:dmm031435 © 2018. Published by The Company of Biologists Ltd