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a549 cells expressing cfp h2b  (Nikon)


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    Nikon a549 cells expressing cfp h2b
    MCRS1 S65 phosphorylation-deficient mutant exhibits a defect in chromosome segregation. (A) Time-lapse live-cell images. <t>A549</t> cells expressing H2B-CFP were transfected with MCRS1 siRNA. Cells were then arrested at G2 by RO3306 and released into fresh medium for monitoring chromosome segregation. Time after nuclear envelope breakdown was counted by minutes. Misaligned chromosomes in MCRS1 siRNA cells were magnified in a rectangle. Bar: 5 μm. (B) Mitotic duration of MCRS1 depleted cells. Results from three independent experiments of panel A were combined and are shown together as mean ± SEM (total 120 mitotic cells). Two-tailed P value was calculated by unpaired Student’s t test. **** means P value is less than 0.0001, *** less than 0.001, and ** less than 0.01. NS stands for not significant. (C) Immunofluorescence of mitotic cells. HeLa Tet-on cells stably expressing MCRS1-GFP were fixed and processed for immunofluorescence using GFP antibody. DNA was stained by Hoechst 33342. Bar: 5 μm. (D) Stable cell line expressing MCRS1-GFP WT, S65A, or S65D. HeLa Tet-on cells were transfected with MCRS1 siRNA and processed for Western blot to examine MCRS1 protein level. (E) Mitotic duration of stable cell lines. Stable cell lines were transfected with siRNA and mitotic duration was counted by live-cell DIC microscope movie. Results from two independent experiments counting ∼40 cells were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. (F) Abnormal chromosome alignment at metaphase. Stable cell lines were transfected with siRNA and treated with MG132 for 1 h. Metaphase cells were counted either normal or abnormal, based on DNA and mitotic spindle distribution. Results from three independent experiments counting ∼60 cells each were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. Bar: 5 μm.
    A549 Cells Expressing Cfp H2b, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 59909 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a549 cells expressing cfp h2b - by Bioz Stars, 2026-10
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    1) Product Images from "Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment"

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment

    Journal: Molecular Biology of the Cell

    doi: 10.1091/mbc.E18-09-0546

    MCRS1 S65 phosphorylation-deficient mutant exhibits a defect in chromosome segregation. (A) Time-lapse live-cell images. A549 cells expressing H2B-CFP were transfected with MCRS1 siRNA. Cells were then arrested at G2 by RO3306 and released into fresh medium for monitoring chromosome segregation. Time after nuclear envelope breakdown was counted by minutes. Misaligned chromosomes in MCRS1 siRNA cells were magnified in a rectangle. Bar: 5 μm. (B) Mitotic duration of MCRS1 depleted cells. Results from three independent experiments of panel A were combined and are shown together as mean ± SEM (total 120 mitotic cells). Two-tailed P value was calculated by unpaired Student’s t test. **** means P value is less than 0.0001, *** less than 0.001, and ** less than 0.01. NS stands for not significant. (C) Immunofluorescence of mitotic cells. HeLa Tet-on cells stably expressing MCRS1-GFP were fixed and processed for immunofluorescence using GFP antibody. DNA was stained by Hoechst 33342. Bar: 5 μm. (D) Stable cell line expressing MCRS1-GFP WT, S65A, or S65D. HeLa Tet-on cells were transfected with MCRS1 siRNA and processed for Western blot to examine MCRS1 protein level. (E) Mitotic duration of stable cell lines. Stable cell lines were transfected with siRNA and mitotic duration was counted by live-cell DIC microscope movie. Results from two independent experiments counting ∼40 cells were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. (F) Abnormal chromosome alignment at metaphase. Stable cell lines were transfected with siRNA and treated with MG132 for 1 h. Metaphase cells were counted either normal or abnormal, based on DNA and mitotic spindle distribution. Results from three independent experiments counting ∼60 cells each were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. Bar: 5 μm.
    Figure Legend Snippet: MCRS1 S65 phosphorylation-deficient mutant exhibits a defect in chromosome segregation. (A) Time-lapse live-cell images. A549 cells expressing H2B-CFP were transfected with MCRS1 siRNA. Cells were then arrested at G2 by RO3306 and released into fresh medium for monitoring chromosome segregation. Time after nuclear envelope breakdown was counted by minutes. Misaligned chromosomes in MCRS1 siRNA cells were magnified in a rectangle. Bar: 5 μm. (B) Mitotic duration of MCRS1 depleted cells. Results from three independent experiments of panel A were combined and are shown together as mean ± SEM (total 120 mitotic cells). Two-tailed P value was calculated by unpaired Student’s t test. **** means P value is less than 0.0001, *** less than 0.001, and ** less than 0.01. NS stands for not significant. (C) Immunofluorescence of mitotic cells. HeLa Tet-on cells stably expressing MCRS1-GFP were fixed and processed for immunofluorescence using GFP antibody. DNA was stained by Hoechst 33342. Bar: 5 μm. (D) Stable cell line expressing MCRS1-GFP WT, S65A, or S65D. HeLa Tet-on cells were transfected with MCRS1 siRNA and processed for Western blot to examine MCRS1 protein level. (E) Mitotic duration of stable cell lines. Stable cell lines were transfected with siRNA and mitotic duration was counted by live-cell DIC microscope movie. Results from two independent experiments counting ∼40 cells were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. (F) Abnormal chromosome alignment at metaphase. Stable cell lines were transfected with siRNA and treated with MG132 for 1 h. Metaphase cells were counted either normal or abnormal, based on DNA and mitotic spindle distribution. Results from three independent experiments counting ∼60 cells each were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. Bar: 5 μm.

    Techniques Used: Phospho-proteomics, Mutagenesis, Expressing, Transfection, Two Tailed Test, Immunofluorescence, Stable Transfection, Staining, Western Blot, Microscopy

    Related Articles

    Fluorescence:

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B was recorded every 10 min for a total duration of 24 h with a 10× objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO 2 -controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B were recorded every 10 min for a total duration of 24 h with a 10X objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO2-controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Imaging:

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B was recorded every 10 min for a total duration of 24 h with a 10× objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO 2 -controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B were recorded every 10 min for a total duration of 24 h with a 10X objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO2-controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Expressing:

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B was recorded every 10 min for a total duration of 24 h with a 10× objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO 2 -controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B were recorded every 10 min for a total duration of 24 h with a 10X objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO2-controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Microscopy:

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B was recorded every 10 min for a total duration of 24 h with a 10× objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO 2 -controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B were recorded every 10 min for a total duration of 24 h with a 10X objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO2-controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Incubation:

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B was recorded every 10 min for a total duration of 24 h with a 10× objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO 2 -controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment
    Article Snippet: Law images from Nikon microscope were deconvoluted by NIS-element (Nikon) software and processed with ImageJ and Photoshop (Adobe). .. Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B were recorded every 10 min for a total duration of 24 h with a 10X objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO2-controlled stage incubation unit (Okolab). .. MCRS1 proteins were purified from HeLa Tet-on cells stably expressing MCRS1-GFP-Strep by Strep-Tactin beads (IBA Lifescience) and digested by LysC enzyme (Promega).



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    a549 cells expressing cfp h2b  (Nikon)
    99
    Nikon a549 cells expressing cfp h2b
    MCRS1 S65 phosphorylation-deficient mutant exhibits a defect in chromosome segregation. (A) Time-lapse live-cell images. <t>A549</t> cells expressing H2B-CFP were transfected with MCRS1 siRNA. Cells were then arrested at G2 by RO3306 and released into fresh medium for monitoring chromosome segregation. Time after nuclear envelope breakdown was counted by minutes. Misaligned chromosomes in MCRS1 siRNA cells were magnified in a rectangle. Bar: 5 μm. (B) Mitotic duration of MCRS1 depleted cells. Results from three independent experiments of panel A were combined and are shown together as mean ± SEM (total 120 mitotic cells). Two-tailed P value was calculated by unpaired Student’s t test. **** means P value is less than 0.0001, *** less than 0.001, and ** less than 0.01. NS stands for not significant. (C) Immunofluorescence of mitotic cells. HeLa Tet-on cells stably expressing MCRS1-GFP were fixed and processed for immunofluorescence using GFP antibody. DNA was stained by Hoechst 33342. Bar: 5 μm. (D) Stable cell line expressing MCRS1-GFP WT, S65A, or S65D. HeLa Tet-on cells were transfected with MCRS1 siRNA and processed for Western blot to examine MCRS1 protein level. (E) Mitotic duration of stable cell lines. Stable cell lines were transfected with siRNA and mitotic duration was counted by live-cell DIC microscope movie. Results from two independent experiments counting ∼40 cells were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. (F) Abnormal chromosome alignment at metaphase. Stable cell lines were transfected with siRNA and treated with MG132 for 1 h. Metaphase cells were counted either normal or abnormal, based on DNA and mitotic spindle distribution. Results from three independent experiments counting ∼60 cells each were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. Bar: 5 μm.
    A549 Cells Expressing Cfp H2b, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/a549+cells+expressing+cfp+h2b/Objectives/pmc06724509-174-4-26
    Average 99 stars, based on 1 article reviews
    a549 cells expressing cfp h2b - by Bioz Stars, 2026-10
    99/100 stars
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    MCRS1 S65 phosphorylation-deficient mutant exhibits a defect in chromosome segregation. (A) Time-lapse live-cell images. A549 cells expressing H2B-CFP were transfected with MCRS1 siRNA. Cells were then arrested at G2 by RO3306 and released into fresh medium for monitoring chromosome segregation. Time after nuclear envelope breakdown was counted by minutes. Misaligned chromosomes in MCRS1 siRNA cells were magnified in a rectangle. Bar: 5 μm. (B) Mitotic duration of MCRS1 depleted cells. Results from three independent experiments of panel A were combined and are shown together as mean ± SEM (total 120 mitotic cells). Two-tailed P value was calculated by unpaired Student’s t test. **** means P value is less than 0.0001, *** less than 0.001, and ** less than 0.01. NS stands for not significant. (C) Immunofluorescence of mitotic cells. HeLa Tet-on cells stably expressing MCRS1-GFP were fixed and processed for immunofluorescence using GFP antibody. DNA was stained by Hoechst 33342. Bar: 5 μm. (D) Stable cell line expressing MCRS1-GFP WT, S65A, or S65D. HeLa Tet-on cells were transfected with MCRS1 siRNA and processed for Western blot to examine MCRS1 protein level. (E) Mitotic duration of stable cell lines. Stable cell lines were transfected with siRNA and mitotic duration was counted by live-cell DIC microscope movie. Results from two independent experiments counting ∼40 cells were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. (F) Abnormal chromosome alignment at metaphase. Stable cell lines were transfected with siRNA and treated with MG132 for 1 h. Metaphase cells were counted either normal or abnormal, based on DNA and mitotic spindle distribution. Results from three independent experiments counting ∼60 cells each were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. Bar: 5 μm.

    Journal: Molecular Biology of the Cell

    Article Title: Mps1 regulates spindle morphology through MCRS1 to promote chromosome alignment

    doi: 10.1091/mbc.E18-09-0546

    Figure Lengend Snippet: MCRS1 S65 phosphorylation-deficient mutant exhibits a defect in chromosome segregation. (A) Time-lapse live-cell images. A549 cells expressing H2B-CFP were transfected with MCRS1 siRNA. Cells were then arrested at G2 by RO3306 and released into fresh medium for monitoring chromosome segregation. Time after nuclear envelope breakdown was counted by minutes. Misaligned chromosomes in MCRS1 siRNA cells were magnified in a rectangle. Bar: 5 μm. (B) Mitotic duration of MCRS1 depleted cells. Results from three independent experiments of panel A were combined and are shown together as mean ± SEM (total 120 mitotic cells). Two-tailed P value was calculated by unpaired Student’s t test. **** means P value is less than 0.0001, *** less than 0.001, and ** less than 0.01. NS stands for not significant. (C) Immunofluorescence of mitotic cells. HeLa Tet-on cells stably expressing MCRS1-GFP were fixed and processed for immunofluorescence using GFP antibody. DNA was stained by Hoechst 33342. Bar: 5 μm. (D) Stable cell line expressing MCRS1-GFP WT, S65A, or S65D. HeLa Tet-on cells were transfected with MCRS1 siRNA and processed for Western blot to examine MCRS1 protein level. (E) Mitotic duration of stable cell lines. Stable cell lines were transfected with siRNA and mitotic duration was counted by live-cell DIC microscope movie. Results from two independent experiments counting ∼40 cells were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. (F) Abnormal chromosome alignment at metaphase. Stable cell lines were transfected with siRNA and treated with MG132 for 1 h. Metaphase cells were counted either normal or abnormal, based on DNA and mitotic spindle distribution. Results from three independent experiments counting ∼60 cells each were combined and are shown here as mean ± SEM. Two-tailed P value was calculated similarly as in panel B. Bar: 5 μm.

    Article Snippet: Fluorescence time-lapse imaging of A549 cells expressing CFP-H2B was recorded every 10 min for a total duration of 24 h with a 10× objective in a Nikon Eclipse Ti-E microscope equipped with a temperature- and CO 2 -controlled stage incubation unit (Okolab).

    Techniques: Phospho-proteomics, Mutagenesis, Expressing, Transfection, Two Tailed Test, Immunofluorescence, Stable Transfection, Staining, Western Blot, Microscopy