Fig. 3: Nuclear export signal (NES) of ITSN1-S located within its SH3 domains. | Cell Death & Disease

Fig. 3: Nuclear export signal (NES) of ITSN1-S located within its SH3 domains.

From: Endocytic protein intersectin1-S shuttles into nucleus to suppress the DNA replication in breast cancer

Fig. 3

A Representative immunofluorescence images of MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h (+LMB) or without LMB as a control (−LMB). Localization of EH domains (EH1,2) of ITSN1-S was detected using anti-flag and anti-HA antibodies, respectively. Scale bars, 25 μm. Quantitative results were analyzed in the right panel. Values were expressed as mean ± SD from three independent experiments (two-tailed Student’s t-test). ns, no significance. B Expression of EH domains of ITSN1-S in the cytoplasm (Cyto) and nuclei (Nuc) was detected by Western blot analysis in MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h or without LMB as a control. β-actin and histone were used as specific markers for cytoplasm and nuclei, respectively. C Representative immunofluorescence images of MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h (+LMB) or without LMB as a control (−LMB). Localization of the CC domain of ITSN1-S was detected using anti-flag and anti-HA antibodies, respectively. Scale bars, 25 μm. Quantitative results were analyzed in the right panel. Values were expressed as mean ± SD from three independent experiments (two-tailed Student’s t-test). ns, no significance. D Expression of CC domain of ITSN1-S in the cytoplasm (Cyto) and nuclei (Nuc) was detected by Western blot analysis in MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h or without LMB as a control. β-actin and histone were used as specific markers for cytoplasm and nuclei, respectively. E Representative immunofluorescence images of MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h (+LMB) or without LMB as a control (−LMB). Localization of SH3 domains (5SH3) of ITSN1-S was detected using anti-flag and anti-HA antibodies, respectively. Scale bars, 25 μm. Quantitative results were analyzed in the right panel. Values were expressed as mean ± SD from three independent experiments (two-tailed Student’s t-test, ***P < 0.001). F Expression of SH3 domains of ITSN1-S in the cytoplasm (Cyto) and nuclei (Nuc) was detected by Western blot analysis in MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h or without LMB as a control. β-actin and histone were used as specific markers for cytoplasm and nuclei, respectively. G Representative immunofluorescence images of KOITSN1/MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h (+LMB) or without LMB as a control (−LMB). Localization of EH domains (EH1,2) of ITSN1-S was detected using anti-flag and anti-HA antibodies, respectively. Scale bars, 25 μm. Quantitative results were analyzed in the right panel. Values were expressed as mean ± SD from three independent experiments (two-tailed Student’s t-test). ns, no significance. H Representative immunofluorescence images of KOITSN1/MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h (+LMB) or without LMB as a control (−LMB). Localization of CC domains (CC) of ITSN1-S was detected using anti-flag and anti-HA antibodies, respectively. Scale bars, 25 μm. Quantitative results were analyzed in the right panel. Values were expressed as mean ± SD from three independent experiments (two-tailed Student’s t-test). ns, no significance. I Representative immunofluorescence images of KOITSN1/MDA-MB-231 cells treated with 10 ng/ml LMB for 5 h (+LMB) or without LMB as a control (−LMB). Localization of SH3 domains (5SH3) of ITSN1-S was detected using anti-flag and anti-HA antibodies, respectively. Scale bars, 25 μm. Quantitative results were analyzed in the right panel. Values were expressed as mean ± SD from three independent experiments (two-tailed Student’s t-test, ***P < 0.001).

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