Skip to main content
. 2022 Aug 10;48(6):e12837. doi: 10.1111/nan.12837

FIGURE 2.

FIGURE 2

Acute monoallelic Nbn loss induces DNA damage response (DDR) activation in Smo agonist (SAG)‐dependent cerebellar neurospheres (S‐cNS). (A) Schematic protocol depicting the generation and manipulation of Nbn F6/+;Gli1CreER+/− S‐cNS. (B) Western blot (WB) analysis of the Nbn F6/+;Gli1CreER+/− S‐cNS treated with ethanol (ETOH) or 4‐hydroxytamoxifen (4‐OHT) for 24 h and collected 48 h after the washout. Blots were probed with the indicated antibodies, and β‐actin was used as loading control. c‐PARP, cleaved PARP. Cre‐induced Nbn gene cleavage identified by PCR is provided in the lowest panel, as an internal control; the size of wild type and floxed alleles are indicated as well as the Δ6 deleted allele. Data are representative of four replicates. (C) Representative images of Nbn F6/+;Gli1CreER+/− S‐cNS treated with ETOH or 4‐OHT for 24 h and collected 48 h after the washout. Cells were suspended in a Trypan blue solution to mark dead cells. (D) Representative phase‐contrast images of Nbn F6/+;Gli1CreER+/− adherent S‐cNS treated with ETOH or 4‐OHT for 24 h and photographed 48 h after washout. Arrows highlight dead cells (original magnification: 40×). (E) Quantification of cell death in cells treated as in D. Data are representative of three replicates.