kn92 422709  (Millipore)

Journal: Oncotarget

Article Title: Ca 2+ /calmodulin-dependent protein kinase IIγ enhances stem-like traits and tumorigenicity of lung cancer cells

doi:

Figure Lengend Snippet: A. Relative gene expression of OCT4, NANOG, MYC, and KLF4 in ZRLC-1 or HCC827 cells treated with DMSO, KN92, or KN93. Data are expressed as mean ± SEM of n = 3 independent cell dishes per condition. * p < 0.05, ** p < 0.01, *** p < 0.001 versus cells treated with KN92. B. Detection of Oct4, c-Myc, pCaMKIIγ, and GAPDH protein by western blots in ZRLC-1 or HCC827 cells treated with DMSO, KN92, or KN93. C. Quantitative analysis of oncosphere formation by 3000 ZRLC-1 or HCC827 cells treated with DMSO, KN92, or KN93. Data are expressed as mean ± SEM of n = 3 independent cell dishes per condition. * p < 0.05 versus cells treated with KN92. D. ZRLC-1 or HCC827 cells pre-treated with KN92 or KN93 were separately injected subcutaneously into nude mice. Data are expressed as mean ± SEM of n = 5 mice per group. * p < 0.05, ** p < 0.01. NS: no significance. Tumor incidence is displayed on the graph.

Article Snippet: KN93 (Cat. No. 422711), KN92 (Cat. No. 422709), Akt Inhibitor IV (Cat. No. 124011), β-catenin/Tcf Inhibitor V (Cat. No. 219334), and Wnt Pathway inhibitor XI (Cat. No. 681674) were purchased from Merck Millipore.

Techniques: Expressing, Western Blot, Injection

Journal: Oncotarget

Article Title: Ca 2+ /calmodulin-dependent protein kinase IIγ enhances stem-like traits and tumorigenicity of lung cancer cells

doi:

Figure Lengend Snippet: A. Detection of Akt and β-catenin signals in H1299 parental or oncosphere cells by western blots. B. Detection of Akt and β-catenin signals in H1299 cells tranduced with control or CaMKIIγ vector by western blots. C. Detection of Akt and β-catenin signals in ZRLC-1 cells treated with DMSO, KN92, or KN93 by western blots. D. Detection of Oct4 and c-Myc in CaMKIIγ-overexpressed H1299 cells treated with Akt or β-catenin inhibitor by western blots. E. Lysates of CaMKIIγ-overexpressed A549 and H1299 cells were incubated with CaMKIIγ or Akt antibody for immunoprecipitation and the immune complex was then purified, separated by SDS-PAGE, and immunoblotted with Akt or CaMKIIγ antibody. F. Dual luciferase reporter assay of Wnt pathway (TOP, FOP, and the ratio of TOP/FOP) in control or CaMKIIγ-overexpressed H1299 cells. Data are expressed as mean ± SEM of n = 4 independent cell dishes per condition. ** p < 0.01. G. Quantitative western blot analysis of oncosphere formation by control or CaMKIIγ-overexpressed A549 or H1299 cells treated with Akt or β-catenin inhibitor. Data are expressed as mean ± SEM of n = 4 independent cell dishes per condition. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: KN93 (Cat. No. 422711), KN92 (Cat. No. 422709), Akt Inhibitor IV (Cat. No. 124011), β-catenin/Tcf Inhibitor V (Cat. No. 219334), and Wnt Pathway inhibitor XI (Cat. No. 681674) were purchased from Merck Millipore.

Techniques: Western Blot, Plasmid Preparation, Incubation, Immunoprecipitation, Purification, SDS Page, Luciferase, Reporter Assay

Journal: Oncotarget

Article Title: Ca 2+ /calmodulin-dependent protein kinase IIγ enhances stem-like traits and tumorigenicity of lung cancer cells

doi:

Figure Lengend Snippet: A. Detection of acetylation and methylation of histone in control or CaMKIIγ-overexpressed H1299 cells by western blots. B. Detection of acetylated histone3 (H3Ac) in ZRLC-1 cells treated with DMSO, KN92, or KN93 by western blots. C. Detection of H3Ac, Oct4, and c-Myc in KN93-treated ZRLC-1 cells cultured with TSA by western blots. D. Detection of H3Ac in control or CaMKIIγ-overexpressed H1299 cells treated with Akt inhibitor by western blots. E. CHIP-enriched DNA from control or CaMKIIγ-overexpressed H1299 cells treated with Akt inhibitor using anti-H3Ac antibody is amplified by real-time PCR, depending on primers designed according to the transcription regulatory region of OCT4. Data are expressed as enrichment assessed relative to the input DNA ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. F. Detection of histone acetylation transferases in control or CaMKIIγ-overexpressed H1299 cells by western blots. G. Detection of histone acetylation transferases in H1299 parental or oncosphere cells by western blots. (H) CHIP-enriched DNA from control or CaMKIIγ-overexpressed H1299 cells treated with Akt inhibitor using anti-PCAF antibody is amplified by real-time PCR, depending on primers designed according to the transcription regulatory region of OCT4. Data are expressed as enrichment assessed relative to the input DNA ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: KN93 (Cat. No. 422711), KN92 (Cat. No. 422709), Akt Inhibitor IV (Cat. No. 124011), β-catenin/Tcf Inhibitor V (Cat. No. 219334), and Wnt Pathway inhibitor XI (Cat. No. 681674) were purchased from Merck Millipore.

Techniques: Methylation, Western Blot, Cell Culture, Amplification, Real-time Polymerase Chain Reaction