Figure 7. LIN37 function in DNA end protection is restricted to G0.
(A, B) Western blot analysis of indicated proteins in cycling and non-cycling abl pre-B cells (A) or MCF10A cells (B). (C, D) Western blot analysis of indicated proteins in cycling G1 or S/G2/M abl pre-B cells (C) or MCF10A cells (D), isolated by flow cytometric cell sorting based on the PIP-FUCCI reporter. Representative of two independent experiments. Asterisk indicates non-specific recognizing bands. (E, F) Flow cytometric analysis of chromatin-bound RPA and γH2AX before and after IR treatment of G1-phase Lig4−/−, Lig4−/−:Trp53bp1−/− and Lig4−/−:Lin37−/− abl pre-B cells (E) or WT, Trp53bp1−/−, and Lin37−/− MCF10A cells (F). Representative of three experiments. IR, ionizing radiation; WT, wild type.
Figure 7—source data 1. RNA-Seq result and GO analysis in cycling G1
Lig4−/− and Lig4−/−:Lin37−/− abl pre-B cells.
GO, gene ontology; RNA-Seq, RNA sequencing.
elife-68466-fig7-data1.xlsx (4.8MB, xlsx)
Figure 7—source data 2. GO analysis of genes upregulated in non-cycling G0 and/or cycling G1
Lig4−/−:Lin37−/− abl pre-B cells.
GO, gene ontology.
elife-68466-fig7-data2.xlsx (47.9KB, xlsx)
Figure 7—figure supplement 1. Identification of G1-phase cells in proliferating cells.
(A, B) Flow cytometric analysis of EdU incorporation and DNA content (7-AAD) of cycling Lig4−/−, Lig4−/−:Trp53bp1−/−, and Lig4−/−:Lin37−/− abl pre-B cells (A) or cycling WT, Trp53bp1−/−, and Lin37−/− MCF10A cells (B). Cells were treated (+EdU) or not treated (−EdU) with EdU and analyses were carried out before or after IR. The percentage of G1-phase cells is shown. IR, ionizing radiation; WT, wild type.