Research Paper Volume 13, Issue 7 pp 9874—9899

DNASE1L3 arrests tumor angiogenesis by impairing the senescence-associated secretory phenotype in response to stress

DNASE1L3 impairs angiogenesis by interacting with H2BE. (A) qPCR analysis of chromosomal DNA in the cytoplasmic fraction of differently treated HepG2 cells. (B) Representative images of SA-β-Gal staining of cells in differently treated groups (scale bar, 100 μm). (C) Statistics of SA-β-Gal staining cells in differently treated groups. (D) Immunoblot analysis of inducible expression change of senescence associated signal pathway and downstream proteins including p53, p65, SPINK1 and AREG in differently treated groups. (E, F) The motility of HUVECs were assessed by wound healing assay, the supernatants from cells in different treated groups were added into the culture of HUVECs, images were taken at 0h and 24h (scale bar, 100 μm). (G, H) The cellular migration ability of HUVECs were determined by the transwell migration assay. Cell supernatants from in differently treated groups were added into the lower chamber, images were taken after 24h of incubation (scale bar, 100 μm). (I, J) The tube formation ability of HUVECs were determined by tube formation assay. Cell supernatants from differently treated groups were added into the culture, images were taken after 6h of incubation (scale bar, 100 μm). The results show the means ± SD from at least three separate experiments.

Figure 6. DNASE1L3 impairs angiogenesis by interacting with H2BE. (A) qPCR analysis of chromosomal DNA in the cytoplasmic fraction of differently treated HepG2 cells. (B) Representative images of SA-β-Gal staining of cells in differently treated groups (scale bar, 100 μm). (C) Statistics of SA-β-Gal staining cells in differently treated groups. (D) Immunoblot analysis of inducible expression change of senescence associated signal pathway and downstream proteins including p53, p65, SPINK1 and AREG in differently treated groups. (E, F) The motility of HUVECs were assessed by wound healing assay, the supernatants from cells in different treated groups were added into the culture of HUVECs, images were taken at 0h and 24h (scale bar, 100 μm). (G, H) The cellular migration ability of HUVECs were determined by the transwell migration assay. Cell supernatants from in differently treated groups were added into the lower chamber, images were taken after 24h of incubation (scale bar, 100 μm). (I, J) The tube formation ability of HUVECs were determined by tube formation assay. Cell supernatants from differently treated groups were added into the culture, images were taken after 6h of incubation (scale bar, 100 μm). The results show the means ± SD from at least three separate experiments.