Fig. 3: MARVELD1 binds with PARP1 and clusters in the nucleus during HU stress. | Cell Death & Differentiation

Fig. 3: MARVELD1 binds with PARP1 and clusters in the nucleus during HU stress.

From: The regulation loop of MARVELD1 interacting with PARP1 in DNA damage response maintains genome stability and promotes therapy resistance of cancer cells

Fig. 3

a PPI were conducted from mass spectrometry data (Fig. 2) by the common interactors of PARP1 and MARVELD1. b The interaction of MARVELD1 and PARP1 was analyzed by IP with IgG or the indicated antibody after treated with 8 mM HU (HeLa cells) or 4 mM HU (HEK293T cells) for 24 h, followed by WB. c Schematic diagram showing the domains of PARP1. ZF zinc finger, NLS nuclear localization signal, BRCT BRCA1 C-terminus, WGR Trp-Gly-Arg domain, HD helical subdomain, ART ADP-ribosyl transferase. Truncated PARP1-flag and V5-tagged MARVELD1 were co-transfected into HeLa and HEK293T cells. Lysates were analyzed by IP of Flag with a subsequent WB. d Nuclear and cytoplasmic protein extraction assays of HeLa cells with the indicated treatment for 24 h were conducted, followed by WB. e IF staining of HeLa cells was used to determine the nucleus translocation of MARVELD1 after HU treatment, and the line graphs represent the fluorescence intensity of MARVELD1 at the arrow by ZEN software. Scale bar: 10 μm. f MARVELD1 clustered in nucleus was analyzed by IF staining when HeLa cells were treated by different dose of HU, CPT or Aph. Green staining indicates MARVELD1, blue staining indicates DAPI. The relative nuclear MARVELD1 amount (integrated density/area) per cell was quantified by ImageJ and 50 cells were measured per treatment. Scale bar: 10 μm. ***p < 0.001.

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