ikk alpha antibody (14a231) - bsa free  (Bio-Techne corporation)

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: IKKα-STAT3-S727 axis: a novel mechanism in DOX-induced cardiomyopathy

doi: 10.1007/s00018-024-05439-1

Figure Lengend Snippet: Increased expression of IKKα in mononuclear cells and macrophages in DCM. A Mononuclear cells from the peripheral blood of patients with tumours treated with DOX and healthy controls were purified. Representative flow cytometry plot of the F4/80 and CD11c levels in mononuclear cells; n = 5 per group. B mRNA levels of IKKα in mononuclear cells from patients with tumours treated with DOX and healthy controls. The transcripts levels were normalized to GAPDH. C Representative immunostaining images for macrophages (CD68, red), IKKα (green), and DAPI (blue) in the heart tissues of dilated cardiomyopathy patients. Magnification 600X. D Representative Western blot analysis of IKKα levels in the heart tissue of mice treated with or without DOX was performed with GAPDH as a loading control; N = 6 in each group. E Representative immunostaining images for macrophages (CD68, green), IKKα (red), and DAPI (blue) in the heart tissues of C57BL/6 mice treated with or without DOX. Magnification 600X. * P < 0.05, ** P < 0.01. P values were calculated using the unpaired Student’s t-test. Quantification of macrophages colocalized with IKKα

Article Snippet: The sections were then blocked with goat serum (ZLI-9022, Beijing Zhongshan Biotechnology) for 1 h. Following the blocking step, the sections were incubated overnight at 4 °C with the following primary antibodies: IKKα (NB100-56704, Novus), CD68 (ab53444, Abcam), STAT3 (ab119352, Abcam), and iNOS (ab178945, Abcam).

Techniques: Expressing, Purification, Flow Cytometry, Immunostaining, Western Blot, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: IKKα-STAT3-S727 axis: a novel mechanism in DOX-induced cardiomyopathy

doi: 10.1007/s00018-024-05439-1

Figure Lengend Snippet: IKKα deficiency aggravated cardiac oxidative stress and macrophage reprogramming induced by DOX. A Representative immunostaining images for macrophages (CD68, green), proinflammatory macrophages (iNOS, red), and DAPI (blue) in the heart tissues of IKKα flox/flox and mIKKα −/− mice. Magnification 400X. B mRNA levels of TNF-α, IL-6 and IL-1β in the heart tissues of IKKα flox/flox and mIKKα −/− mice. The transcript levels were normalized to those of GAPDH. C Representative immunostaining images for DHE and TUNEL in the heart tissues of IKKα flox/flox and mIKKα −/− mice. Magnification 100X. D Representative images of heart tissues from IKKα flox/flox and mIKKα −/− mice stained with 4-HNE. Magnification 400X. E Serum levels of LDH, MDA, GSH and SOD in IKKα flox/flox and mIKKα −/− mice; N = 6 in each group. F Metabolism analysis of the OCR and ECAR in macrophages purified from IKKα flox/flox and mIKKα −/− mice treated with DOX. Quantification of macrophage oxidative phosphorylation and glycolysis as determined by the OCR and ECAR. ** P < 0.05, ** P < 0.01, ** P < 0.001, ns: not significant. P values were calculated using an unpaired Student’s t-test

Article Snippet: The sections were then blocked with goat serum (ZLI-9022, Beijing Zhongshan Biotechnology) for 1 h. Following the blocking step, the sections were incubated overnight at 4 °C with the following primary antibodies: IKKα (NB100-56704, Novus), CD68 (ab53444, Abcam), STAT3 (ab119352, Abcam), and iNOS (ab178945, Abcam).

Techniques: Immunostaining, TUNEL Assay, Staining, Purification, Phospho-proteomics

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: IKKα-STAT3-S727 axis: a novel mechanism in DOX-induced cardiomyopathy

doi: 10.1007/s00018-024-05439-1

Figure Lengend Snippet: STAT3 is a marker of a DOX-related proinflammatory macrophage cluster. IKKα flox/flox and macrophage IKKα knockout (mIKKα −/− ) mice were treated with or without DOX for 2 weeks. A Signature of the proinflammatory process for each cluster. The signature score was calculated on the basis of proinflammatory factor score (STAT3, IL6Ra, CCL5, TNFAIP3, TGFB1, IL-1β, TNF-α, HMGB1, CCL2, Hmox1, CCL4, NFKB1, TLR2, CD14, NFE2L2, PLA2G7, NLRP3, CXCL2, JUN, and CCL3). B and C GO analysis (B) and KEGG analysis (C) of macrophage cluster 3. D Heatmap of genes associated with macrophage cluster 3 in IKKα flox/flox and mIKKα −/− mice. E Pearson correlation coefficients between the mRNA levels of STAT3 and representative cytokines, and chemokines. F GO analysis of STAT3hi macrophages in cluster 3

Article Snippet: The sections were then blocked with goat serum (ZLI-9022, Beijing Zhongshan Biotechnology) for 1 h. Following the blocking step, the sections were incubated overnight at 4 °C with the following primary antibodies: IKKα (NB100-56704, Novus), CD68 (ab53444, Abcam), STAT3 (ab119352, Abcam), and iNOS (ab178945, Abcam).

Techniques: Marker, Knock-Out

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: IKKα-STAT3-S727 axis: a novel mechanism in DOX-induced cardiomyopathy

doi: 10.1007/s00018-024-05439-1

Figure Lengend Snippet: IKKα colocalized with STAT3 and increased STAT3-S727 phosphorylation. A . Representative immunostaining images for macrophages (CD68, red), IKKα (red), and STAT3 (purple) in the heart tissues of IKKα flox/flox and mIKKα −/− mice. Magnification 600X. Quantification of STAT3 colocalized with IKKα and macrophages. B .Co-IP analysis of IKKα and STAT3 in primary macrophages from IKKα flox/flox mice stimulated with LPS. C and D Western blotting analysis was used to determine the levels of STAT3, STAT3-Y705 and STAT3-S727 in primary macrophages from IKKα flox/flox and mIKKα −/− mice stimulated with LPS; N = 6 in each group. * P < 0.05, ** P < 0.01, ns: not significant. P values were calculated using an unpaired Student’s t-test

Article Snippet: The sections were then blocked with goat serum (ZLI-9022, Beijing Zhongshan Biotechnology) for 1 h. Following the blocking step, the sections were incubated overnight at 4 °C with the following primary antibodies: IKKα (NB100-56704, Novus), CD68 (ab53444, Abcam), STAT3 (ab119352, Abcam), and iNOS (ab178945, Abcam).

Techniques: Phospho-proteomics, Immunostaining, Co-Immunoprecipitation Assay, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: IKKα-STAT3-S727 axis: a novel mechanism in DOX-induced cardiomyopathy

doi: 10.1007/s00018-024-05439-1

Figure Lengend Snippet: Inhibition of STAT3-S727 alleviated IKKα deficiency-induced cardiac injury and oxidative stress in cardiomyocytes. A and B IKKα flox/flox and macrophage IKKα knockout (mIKKα −/− ) mice induced with DOX were treated with or without niclosamide for 2 weeks. Representative echocardiography images of murine hearts and left ventricular ejection fraction (EF), left ventricular fractional shortening (FS) and left ventricular volume (LV vol) at end-diastole and systole determined by echocardiography; N = 6 in each group. C Western blotting analysis was used to determine the levels of STAT3-S727 in primary macrophages from IKKα flox/flox mice treated with or without niclosamide for 4 h after LPS stimulation. D Representative immunostaining images for DHE in HiPSC-CMs cocultured with primary macrophages stimulated with LPS from IKKα flox/flox and mIKKα −/− mice treated with or without niclosamide for 4 h. Magnification 100X. E Representative immunostaining images for TUNEL in HiPSC-CMs cocultured with primary macrophages stimulated with LPS from IKKα flox/flox and mIKKα −/− mice treated with or without niclosamidefor 4 h. Magnification 400X. * P < 0.05, ** P < 0.01. P values were calculated using an unpaired Student’s t-test

Article Snippet: The sections were then blocked with goat serum (ZLI-9022, Beijing Zhongshan Biotechnology) for 1 h. Following the blocking step, the sections were incubated overnight at 4 °C with the following primary antibodies: IKKα (NB100-56704, Novus), CD68 (ab53444, Abcam), STAT3 (ab119352, Abcam), and iNOS (ab178945, Abcam).

Techniques: Inhibition, Knock-Out, Western Blot, Immunostaining, TUNEL Assay