Fig. 2: NKX6.3 regulates DNA damage response. | Oncogenesis

Fig. 2: NKX6.3 regulates DNA damage response.

From: Depletion of NK6 Homeobox 3 (NKX6.3) causes gastric carcinogenesis through copy number alterations by inducing impairment of DNA replication and repair regulation

Fig. 2

A Representative comet images and the mean of tail moments in HFE-145shCtrl, HFE-145shNKX6.3#1, and HFE-145shNKX6.3#2 cells (n > 10). B Immunofluorescent staining for γH2AX, 53BP1, Rad51, and RPA32 in HFE-145shCtrl, HFE-145shNKX6.3#1, and HFE-145shNKX6.3#2 cells. Nuclear size was measured in HFE-145shCtrl, HFE-145shNKX6.3#1, and HFE-145shNKX6.3#2 cells (lower left; n > 100). Foci of cells were scored positive when at least five γH2AX foci colocalized with 53BP1 (lower right; n > 100). C Immunoblot analysis of Rad50, p95/NBS, MRE11, Xrcc1, PCNA, p-ATR, p-ATM, p-Chk1, p-Chk2, Rad51, Rad52, and γH2AX in NKX6.3-depleted cells. D In immunoprecipitation analysis, depletion of NKX6.3 inhibited the formation of the MRN complex. E DR-GFP is shown along with the HDR product that uses iGFP as the template for nascent DNA synthesis, which results in the restoration of a GFP expression cassette (left). EJ5-GFP is shown along with products of EJ between the distal DSB ends (distal-EJ) that restore the GFP expression cassette (right). F Representative dot-plot images of DR-GFP HFE-145shCtrl, HFE-145shNKX6.3#1, and HFE-145shNKX6.3#2 cells showing HR repair (upper left). Quantification of HR repair (upper right; n = 3). Representative dot-plot images of EJ5-GFP HFE-145shCtrl, HFE-145shNKX6.3#1, and HFE-145shNKX6.3#2 cells showing NHEJ repair (lower left). Quantification of NHEJ repair (lower right; n = 3). H Effects of CDT1 and RPA1 on tail moments in HFE-145shNKX6.3#1 and HFE-145shNKX6.3#2 cells (n > 10). G In immunoblot analysis, knockdown of CDT1 and ectopic expression of RPA1 reduced γH2AX expression in HFE-145shNKX6.3#1 and HFE-145shNKX6.3#2 cells.

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