Anti-PATT1抗体

• WB

Unknown

Western blot - Anti-PATT1 antibody (AB106408)

Lane 1:

Western blot - Anti-PATT1 antibody (ab106408) at 1 µg/mL

Lane 2:

Western blot - Anti-PATT1 antibody (ab106408) at 2 µg/mL

All lanes:

Human thymus tissue lysate at 15 µg

Predicted band size: 27 kDa

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• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-PATT1 antibody (AB106408)

Western Blotting using Anti-PATT1 antibody, ab106408. Publication image from Ju, J. et al., 2017, Nat Commun, 29030587. Legend direct from paper.

Regulation of Slug by NatD is acetyltransferase activity-dependent. a (left) In vitro acetylation assay showing the catalytic activity of NatDδ and wild-type NatD (CPM, counts per minute). Data are mean ± s.d. of three independent experiments; Student’s t-test, **P < 0.01 compared with wild-type NatD. (right) SDS-PAGE analysis of purified recombinant NatDδ and wild-type NatD proteins from E. coli stained by Coomassie brilliant blue (CBB). MW, protein molecular weight markers. b (top) Western blot analysis of an H4 (1–31) peptide from in vitro acetylation assay in the presence of NatDδ or wild-type NatD. (bottom) H4 (1–31) peptide shown by Coomassie blue staining. Blots are representative of three independent experiments. c (top) Autoradiographic image showing products from in vitro acetylation assay using histones as substrates extracted from H1299 cells. Results are representative of three independent experiments. (bottom) Histones shown by Coomassie blue staining. d Quantitative real-time PCR analysis of mRNA levels of Slug, E-cadherin, N-cadherin, and Vimentin normalized to GAPDH in H1299 cells overexpressing NatDδ or wild-type NatD. Data are mean ± s.d. of three independent experiments; Student’s t-test, **P < 0.01 compared with the wild-type NatD. e Western blot analysis of indicated proteins from H1299 cells overexpressing NatDδ or wild-type NatD. GAPDH and histone H4 served as loading controls. Data are representative of three independent experiments. f, g Representative images of the migration (e) and invasion (f) of H1299 cells overexpressing NatDδ or wild-type NatD with transwell assay from three independent experiments (top panel). Cell counts for the corresponding assays of at least four random microscope fields (x100 magnification). Cell migration and invasion are expressed as a percentage of control (bottom panel). Results are shown as mean ± s.d. from three independent experiments. Two-tailed Student’s t-test was used. **P < 0.01 compared with the indicated control

• WB

CiteAb

Western blot - Anti-PATT1 antibody (AB106408)

Western Blotting using Anti-PATT1 antibody, ab106408. Publication image from Ju, J. et al., 2017, Nat Commun, 29030587. Legend direct from paper.

Regulation of Slug by NatD is acetyltransferase activity-dependent. a (left) In vitro acetylation assay showing the catalytic activity of NatDδ and wild-type NatD (CPM, counts per minute). Data are mean ± s.d. of three independent experiments; Student’s t-test, **P < 0.01 compared with wild-type NatD. (right) SDS-PAGE analysis of purified recombinant NatDδ and wild-type NatD proteins from E. coli stained by Coomassie brilliant blue (CBB). MW, protein molecular weight markers. b (top) Western blot analysis of an H4 (1–31) peptide from in vitro acetylation assay in the presence of NatDδ or wild-type NatD. (bottom) H4 (1–31) peptide shown by Coomassie blue staining. Blots are representative of three independent experiments. c (top) Autoradiographic image showing products from in vitro acetylation assay using histones as substrates extracted from H1299 cells. Results are representative of three independent experiments. (bottom) Histones shown by Coomassie blue staining. d Quantitative real-time PCR analysis of mRNA levels of Slug, E-cadherin, N-cadherin, and Vimentin normalized to GAPDH in H1299 cells overexpressing NatDδ or wild-type NatD. Data are mean ± s.d. of three independent experiments; Student’s t-test, **P < 0.01 compared with the wild-type NatD. e Western blot analysis of indicated proteins from H1299 cells overexpressing NatDδ or wild-type NatD. GAPDH and histone H4 served as loading controls. Data are representative of three independent experiments. f, g Representative images of the migration (e) and invasion (f) of H1299 cells overexpressing NatDδ or wild-type NatD with transwell assay from three independent experiments (top panel). Cell counts for the corresponding assays of at least four random microscope fields (x100 magnification). Cell migration and invasion are expressed as a percentage of control (bottom panel). Results are shown as mean ± s.d. from three independent experiments. Two-tailed Student’s t-test was used. **P < 0.01 compared with the indicated control

false

• WB

CiteAb

Western blot - Anti-PATT1 antibody (AB106408)

Western Blotting using Anti-PATT1 antibody, ab106408. Publication image from Ju, J. et al., 2017, Nat Commun, 29030587. Legend direct from paper.

NatD is required for lung cancer cell migration and invasion in vitro. a Quantitative real-time PCR analysis of NatD mRNA levels normalized to GAPDH in scrambled control cells (Scr) and NatD-KD1 and NatD-KD2 cells. Results are shown as mean ± s.d. from three independent experiments. Two-tailed Student’s t-test was used. **P < 0.01 compared to Scr control. b Western blot analysis of NatD and Nt-ac-H4 protein levels in scrambled, NatD-KD1, and NatD-KD2 cells. GAPDH and histone H4 served as loading controls. Blots are representative of three independent experiments. c Representative images from wound healing assay of scrambled, NatD-KD1, and NatD-KD2 cells from three independent experiments (left panels). Wound healing assay results are quantified in the histogram (right panel). Results are shown as mean ± s.d. from three independent experiments. Two-tailed Student’s t-test was used. **P < 0.01 compared to Scr control. d Representative images of the migration of scrambled, NatD-KD1, and NatD-KD2 cells in a time-lapse cell tracker migration assay from three independent experiments. Representative images of the migration (e) and invasion (f) of scramble, NatD-KD1, and NatD-KD2 cells with transwell assay from three independent experiments (top panel). Cell counts for the corresponding assays of at least four random microscope fields (x100 magnification). Cell migration and invasion are expressed as a percentage of control (bottom panel). Results are shown as mean ± s.d. from three independent experiments. Two-tailed Student’s t-test was used. **P < 0.01 compared to Scr control

false

• WB

CiteAb

Western blot - Anti-PATT1 antibody (AB106408)

Western Blotting using Anti-PATT1 antibody, ab106408. Publication image from Ju, J. et al., 2017, Nat Commun, 29030587. Legend direct from paper.

Nt-acetylation of histone H4 antagonizes phosphorylation of histone H4 serine 1 to regulate Slug expression. a Western blot analysis of indicated histone H4 modifications in Scr and NatD-KD H1299 cells. Histone H4 served as a loading control. b ChIP analysis of the enrichment of indicated histone H4 modifications on the Slug promoter in Scr and NatD-KD H1299 cells. IgG served as a negative control. Results are shown as mean ± s.d. from three independent experiments; two-tailed Student’s t-test, *P < 0.05, **P < 0.01 compared with the Scr control. c ChIP analysis of the enrichment of H3K4me3 and H3K27me3 on the Slug promoter in Scr and NatD-KD H1299 cells. IgG served as a negative control. Results are shown as mean ± s.d. from three independent experiments; two-tailed Student’s t-test, **P < 0.01 compared with the Scr control. d Western blot analysis of indicated proteins from H1299 cells overexpressing NatDδ or wild-type NatD. Histone H4 served as a loading control. e ChIP analysis of the enrichment of Nt-ac-H4 and H4S1ph on Slug promoter in H1299 cells overexpressing NatDδ or wild-type NatD. IgG served as a negative control. Results are shown as mean ± s.d. from three independent experiments; two-tailed Student’s t-test, *P < 0.05, **P < 0.01 compared with the wild-type control

false

• WB

CiteAb

Western blot - Anti-PATT1 antibody (AB106408)

Western Blotting using Anti-PATT1 antibody, ab106408. Publication image from Ju, J. et al., 2017, Nat Commun, 29030587. Legend direct from paper.

Silencing NatD suppresses cancer cell EMT by downregulating Slug. a Representative phase contrast images of Scr and NatD-KD H1299 cells treated with TGF-β1. Data are representative of three independent experiments. Scale bar, 100 µm. b Quantitative real-time PCR analysis of mRNA levels of indicated key EMT-related genes in Scr and NatD-KD H1299 cells normalized to GAPDH in the absence or presence of TGF-β1. Results are shown as mean ± s.d. of three independent experiments. Two-tailed Student’s t-test was used. **P < 0.01 or *P < 0.05 compared with the indicated control. c Western blot analysis of indicated protein levels in Scr and NatD-KD H1299 cells in the absence or presence of TGF-β1. GAPDH served as a loading control. Data are representative of three independent experiments. d Immunofluorescence analysis of Scr and NatD-KD H1299 cells in the absence or presence of TGF-β1 stained for E-cadherin and N-cadherin. Data are representative of three independent experiments. Scale bar, 20 µm. Migration (e, top) and invasion (f, top) of Scr cells, NatD-KD cells, and NatD-KD cells with enforced Slug expression (NatD-KD + Slug). (bottom panels) Cells were counted in at least four random microscope fields (x100 magnification) for the corresponding assays; migration and invasion are expressed as a percentage of control. Results are shown as mean ± s.d. of three independent experiments. Two-tailed Student’s t-test was used. **P < 0.01 compared with the indicated control. g Quantitative real-time PCR analysis of the mRNA levels of NatD and indicated key EMT-related genes (normalized to GAPDH) in Scr cells, NatD-KD cells, and NatD-KD + Slug cells. Results are shown as mean ± s.d. of three independent experiments. Two-tailed Student’s t-test was used. **P < 0.01 or *P < 0.05 compared with Scr or indicated control. h Western blot analysis of indicated protein levels in Scr cells, NatD-KD cells, and NatD-KD + Slug cells. GAPDH served as a loading control. Data are representative of three independent blots. i Pearson correlation scatter plot of the H score of Slug and NatD in human lung carcinoma (n = 147); r = 0.6672, P < 0.0001

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