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    phosphorylated tyr1507 alk  (Cell Signaling Technology Inc)


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    anti phospho alk rabbit monoclonal antibody  (Cell Signaling Technology Inc)


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    phospho alk y1507  (Cell Signaling Technology Inc)


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    p alk  (Cell Signaling Technology Inc)


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    anti palk y1507  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti palk y1507
    ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using <t>anti-pALK-Y1507</t> and anti-ALK antibodies. Lamin B was incorporated as a loading control.
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    1) Product Images from "ALK Inhibitors-Induced M Phase Delay Contributes to the Suppression of Cell Proliferation"

    Article Title: ALK Inhibitors-Induced M Phase Delay Contributes to the Suppression of Cell Proliferation

    Journal: Cancers

    doi: 10.3390/cancers12041054

    ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using anti-pALK-Y1507 and anti-ALK antibodies. Lamin B was incorporated as a loading control.
    Figure Legend Snippet: ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using anti-pALK-Y1507 and anti-ALK antibodies. Lamin B was incorporated as a loading control.

    Techniques Used: Western Blot, Staining, Microscopy

    phospho alk tyr1507  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho alk tyr1507
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    rabbit phospho y1507 alk  (Cell Signaling Technology Inc)


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    phospho alk tyr1507  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho alk tyr1507
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    rabbit anti human palk  (Cell Signaling Technology Inc)


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    phospho alk tyr1507  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho alk tyr1507
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    Cell Signaling Technology Inc phosphorylated tyr1507 alk
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    ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using <t>anti-pALK-Y1507</t> and anti-ALK antibodies. Lamin B was incorporated as a loading control.
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    ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using <t>anti-pALK-Y1507</t> and anti-ALK antibodies. Lamin B was incorporated as a loading control.
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    ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using <t>anti-pALK-Y1507</t> and anti-ALK antibodies. Lamin B was incorporated as a loading control.
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    ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using <t>anti-pALK-Y1507</t> and anti-ALK antibodies. Lamin B was incorporated as a loading control.
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    Image Search Results


    ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using anti-pALK-Y1507 and anti-ALK antibodies. Lamin B was incorporated as a loading control.

    Journal: Cancers

    Article Title: ALK Inhibitors-Induced M Phase Delay Contributes to the Suppression of Cell Proliferation

    doi: 10.3390/cancers12041054

    Figure Lengend Snippet: ALK inhibitors delay M phase progression. ( A ) Whole cell lysates were analyzed by western blotting with anti-ALK and anti-Lamin B (loading control) antibodies. ( B ) SH-SY5Y cells were treated with inhibitors for 2 days, and viable cells were determined by WST-8 assay. Relative values of absorbance to solvent control (dimethyl sulfoxide, DMSO) are shown as mean ± SD, calculated from three independent experiments. ( C ) SH-SY5Y cells were synchronized with 4 µM RO-3306 for 20 h. After release, cells were treated with 1 µM crizotinib, 0.5 µM ceritinib, and 0.3 µM TAE684 for 1 h and stained for α-tubulin and DNA. Based on microtubule and chromosome morphology, M phase cells were classified into four groups: prophase/prometaphase (P/PM), metaphase (M), anaphase/telophase (A/T), and cytokinesis (Cyto). The percentages of cells of each group (n > 246 in each experiment) and associated mitotic indices ( n > 453 in each experiment) are plotted as the mean ± SD, calculated from three independent experiments. ( D ) The number of cells with misaligned chromosomes was counted under a microscope, and their percentages are plotted as the mean ± SD of three independent experiments ( n > 246 in each experiment). Representative images of cells with misaligned chromosomes are shown; scale bar = 5 µm. The Tukey–Kramer multiple comparison test was used to calculate p values. ** p < 0.01; N.S., not significant. ( E ) SH-SY5Y cells were treated with inhibitors in the same concentrations as in ( C ) for 1 h, and whole cell lysates were analyzed by Western blotting using anti-pALK-Y1507 and anti-ALK antibodies. Lamin B was incorporated as a loading control.

    Article Snippet: The primary antibodies used for immunofluorescence (IF) and WB analyses were rabbit monoclonal anti-ALK (WB, 1:500–1000; IF, 1:200; #3333S, Cell Signaling Technology, Danvers, MA, USA), anti-pALK Y1507 (WB, 1:1000; #14678, Cell Signaling Technology) and anti-c-Met (WB, 1:1000; #8198, Cell Signaling Technology); rat monoclonal anti-α-tubulin (WB, 1:1000; IF, 1:800; MCA78G, Bio-Rad Laboratories, Hercules, CA, USA) and anti-HA (WB, 1:1000; IF, 1:400; 3F10, Roche, Basel, Switzerland); goat anti-Lamin B (WB, 1:200–500; sc-6216, Santa Cruz Biotechnology, Dallas, TX, USA); and mouse monoclonal anti-Actin (WB, 1:2000; A3853, MilliporeSigma, Burlington, MA, USA).

    Techniques: Western Blot, Staining, Microscopy