Anti-RUNX1 / AML1抗体

• WB

Unknown

Western blot - Anti-RUNX1 / AML1 antibody (AB23980)

This antibody recognized three distinct bands of between 48 and 55 kDa in Jurkat nuclear lysate. These may represent distinct isoforms of Runx1 or may represent post-translationally modified forms.

All lanes:

Western blot - Anti-RUNX1 / AML1 antibody (ab23980) at 1 µg/mL

All lanes:

Jurkat nuclear extract lysate (<a href='/products/unavailable/jurkat-nuclear-extract-lysate-ab14844'>ab14844</a>) at 20 µg

Secondary

All lanes:

Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution

Predicted band size: 48 kDa

Observed band size: 48 kDa,52 kDa,55 kDa

false

• WB

CiteAb

Western blot - Anti-RUNX1 / AML1 antibody (AB23980)

Western Blotting using Anti-RUNX1 / AML1 antibody, ab23980. Publication image from Mhawech-Fauceglia, P. et al., 2016, Nat Commun, 26916619. Legend direct from paper.

Upregulation of β-catenin in RUNX1-deficient breast cancer.(a) RUNX1 mRNA expression in the five major breast cancer subtypes in the breast cancer cohort of TCGA20. Expression levels are significantly different between the subtypes (P=6.8e−38 by analysis of variance). Boxes represent the 25% to 75% quartiles, lines within boxes represent the median levels and whiskers represent the non-outlier range. (b) Genes differentially expressed in ER+ tumours with mutant versus wild-type RUNX1 in the breast cancer patient cohort of TCGA20 (Supplementary Data 1) were interrogated using Ingenuity Pathways Analysis (IPA) for annotations related to major developmental signalling pathways. Line graph represents fold enrichment, and statistical significance (bars) was calculated by Fisher's exact test as implemented in the IPA software. (c) IPA analysis was performed as in b for the differentially expressed genes (Supplementary Data 2) in RUNX1-mutant tumours in the breast cancer patient cohort of Ellis et al.18 (d) Top : representative western blot analyses of the indicated proteins in MCF7 and T47D cells expressing either a nonspecific shRNA (shNS) or shRNAs targeting the Runt domain (shRx1RUNT) or the 3′-UTR (shRx13′-UTR) of RUNX1. Bottom : western blots from three independent experiments were scanned using the ImageJ software, and bar graphs represent mean densitometric values (±s.e.m.) for normalized A-β-cat corrected for β-actin. *P<0.05 by t-test. (e) Western blot analysis of total β-catenin in whole-cell extracts (WCE), as well as cytoplasmic (cyt) and nuclear (nuc) fractions of MCF7 cells expressing the shNS or the shRx13′-UTR RNAs.

false

• WB

CiteAb

Western blot - Anti-RUNX1 / AML1 antibody (AB23980)

Western Blotting using Anti-RUNX1 / AML1 antibody, ab23980. Publication image from Mhawech-Fauceglia, P. et al., 2016, Nat Commun, 26916619. Legend direct from paper.

RUNX1 prevents oestrogen-mediated AXIN1 repression.(a) MCF7/shRx1RUNTdox cells were maintained in 10% charcoal-stripped serum for 48 h, treated as indicated for the following 48 h, and AXIN1 mRNA levels were measured by RT–qPCR and corrected for 18S RNA (mean±s.e.m. of three independent experiments). (b) MCF7/shRx1RUNTdox cells in 10% complete serum were treated as indicated for 48 h, and AXIN1 mRNA levels were measured by RT–qPCR and corrected for 18S RNA (mean±s.e.m. of three independent experiments). *P<0.05 by t-test. (c) Scatter plot of the global E2 responsiveness in the presence (y axis) versus absence (x axis) of RUNX1 in MCF7 cells. (d) The indicated ER+ (left) and ER− (right) mammary epithelial cell lines were engineered with the dox-inducible shRx13′-UTR lentiviral vector and treated with dox for 4 days before western blot analysis of the indicated proteins. (e) RT–qPCR results for Axin1 and Runx1 from predominantly ER+ mature luminal (ML) mammary epithelial cells (left) and predominantly ER− luminal progenitor (LP) cells (right) isolated from RUNX1-knockout and control mammary glands as described in the ‘Methods' section.

false

• WB

CiteAb

Western blot - Anti-RUNX1 / AML1 antibody (AB23980)

Western Blotting using Anti-RUNX1 / AML1 antibody, ab23980. Publication image from Mhawech-Fauceglia, P. et al., 2016, Nat Commun, 26916619. Legend direct from paper.

AXIN1 stabilization normalizes β-catenin and partially restores cell cycle control in RUNX1-depleted cells.(a) MCF7 cells constitutively expressing shRx13′-UTR (shRx1) were treated for 36 h with either 5 µM IWR1 or its dimethyl sulphoxide vehicle followed by western blot analysis of the indicated proteins. MCF7 expressing a nonspecific shRNA (shNS) were analysed as a reference control. (b) Cells as in a were treated as indicated for 6 days and their growth rate was calculated based on MTT assays as in Fig. 2b. *P<0.05 by t-test. (c) AXIN1 and P-β-cat levels were assessed 8 h after the release of MCF7/shRx1RUNTdox cells from a G1/S double thymidine block as in Fig. 6h. Dox treatment (to silence RUNX1) initiated along with the release from the first thymidine block and IWR1 treatment (to stabilize AXIN1) initiated 17 h before harvest. (d) MCF7/shRx1RUNTdox cells were treated for 72 h with dox (to silence RUNX1) and 2 nM docetaxel for 48 h (to induce mitotic slippage) as in Fig. 6f,g and IWR1 was added for the last 24 h before FACS analysis. Data are mean±s.e.m. (n=3). *P<0.05 by t-test. (e) Working model for the tumour suppressor function of RUNX1 in ER+ breast cancer, whereby RUNX1 prevents E2-mediated AXIN1 suppression. Mechanisms linking the RUNX1/AXIN1/β–catenin axis to loss of cell cycle control in RUNX1-deficient ER+ mammary epithelial cells remain to be fully elucidated. They entail stimulation of neither LEF/TCF, nor c-MYC, nor CCND1, nor G1/S phase transition, but are associated instead with deregulated mitosis.

false

• WB

CiteAb

Western blot - Anti-RUNX1 / AML1 antibody (AB23980)

Western Blotting using Anti-RUNX1 / AML1 antibody, ab23980. Publication image from Mhawech-Fauceglia, P. et al., 2016, Nat Commun, 26916619. Legend direct from paper.

Upregulation of β-catenin in RUNX1-deficient breast cancer.(a) RUNX1 mRNA expression in the five major breast cancer subtypes in the breast cancer cohort of TCGA20. Expression levels are significantly different between the subtypes (P=6.8e−38 by analysis of variance). Boxes represent the 25% to 75% quartiles, lines within boxes represent the median levels and whiskers represent the non-outlier range. (b) Genes differentially expressed in ER+ tumours with mutant versus wild-type RUNX1 in the breast cancer patient cohort of TCGA20 (Supplementary Data 1) were interrogated using Ingenuity Pathways Analysis (IPA) for annotations related to major developmental signalling pathways. Line graph represents fold enrichment, and statistical significance (bars) was calculated by Fisher's exact test as implemented in the IPA software. (c) IPA analysis was performed as in b for the differentially expressed genes (Supplementary Data 2) in RUNX1-mutant tumours in the breast cancer patient cohort of Ellis et al.18 (d) Top : representative western blot analyses of the indicated proteins in MCF7 and T47D cells expressing either a nonspecific shRNA (shNS) or shRNAs targeting the Runt domain (shRx1RUNT) or the 3′-UTR (shRx13′-UTR) of RUNX1. Bottom : western blots from three independent experiments were scanned using the ImageJ software, and bar graphs represent mean densitometric values (±s.e.m.) for normalized A-β-cat corrected for β-actin. *P<0.05 by t-test. (e) Western blot analysis of total β-catenin in whole-cell extracts (WCE), as well as cytoplasmic (cyt) and nuclear (nuc) fractions of MCF7 cells expressing the shNS or the shRx13′-UTR RNAs.

false