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Fig. 3 | Journal of Experimental & Clinical Cancer Research

Fig. 3

From: Nuclear Transglutaminase 2 interacts with topoisomerase II⍺ to promote DNA damage repair in lung cancer cells

Fig. 3

Nuclear TG2 was recruited to DSB sites. A: Representative images for immunofluorescence staining of TG2 (Red) in the HT-1080 cells with 53BP1-GFP stable expressions at 30 min and 2 h after laser irradiation. Scale bar, 20 μm. B: colocalization of TG2 and 53BP1 was performed by Image Pro Plus 6.0 software. C: Representative images for immunofluorescence staining of TG2 in the same HT-1080 cells with 53BP1-GFP stable expressions after 2 Gy γ-irradiation. Scale bar, 20 μm. Different from Panel A, here, the cells were analyzed in immunofluorescence staining assay of TG2 after extraction of free TG2 with CSK buffer. D: colocalization of TG2 and 53BP1 was performed by Image J software. E: Total Nuclei proteins were isolated and lysed in a no-salt buffer. Afterward, the soluble nuclear proteins (S3) were separated through low speed centrifugation. The chromatin enriched proteins were further isolated from the final pellet of soluble nuclear proteins (P3). Both soluble proteins and chromatin binding proteins were subjected to SDS page assay, and were immune-blotted by TG2, ORC2 and Ku80 antibodies. F: Chromatin enriched proteins were quantified through image J software to detect the levels of DNA binding

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