hs578t  (ATCC)


Bioz Verified Symbol ATCC is a verified supplier
Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    ATCC hs578t
    SNRPD2, SNRPD3 and NHP2L1 protein interactions. a Characterization of <t>Hs578T</t> GFP BAC-reporter cell lines for splicing factors SNRPD2, SNRPD3 and NHP2L1. Images were captured 72 h after transfection. Scale bar = 30 μm. b Overlap of proteins residing in SNRPD2, SNRPD3 and NHP2L1 complexes, respectively. Splicing factors belonging to the nine candidates selected from the primary screen are labeled in purple. Orange: proteins that were selected for further validation. c Nuclear phenotype 72 h after SUN2 knockdown in Hs578T and MDA-MB-231 cells. Scale bar = 30 μm. d Co-immunoprecipitation of SNRPD2, SNRPD3 and NHP2L1 Hs578T BAC GFP-reporters with SUN2. e Alternative splicing of sororin intron 1 upon SUN2 knockdown. Experiments were performed in biological triplicates. Significance was determined using a Student’s T-test. * p
    Hs578t, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hs578t/product/ATCC
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hs578t - by Bioz Stars, 2022-10
    97/100 stars

    Images

    1) Product Images from "Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention"

    Article Title: Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-021-01863-4

    SNRPD2, SNRPD3 and NHP2L1 protein interactions. a Characterization of Hs578T GFP BAC-reporter cell lines for splicing factors SNRPD2, SNRPD3 and NHP2L1. Images were captured 72 h after transfection. Scale bar = 30 μm. b Overlap of proteins residing in SNRPD2, SNRPD3 and NHP2L1 complexes, respectively. Splicing factors belonging to the nine candidates selected from the primary screen are labeled in purple. Orange: proteins that were selected for further validation. c Nuclear phenotype 72 h after SUN2 knockdown in Hs578T and MDA-MB-231 cells. Scale bar = 30 μm. d Co-immunoprecipitation of SNRPD2, SNRPD3 and NHP2L1 Hs578T BAC GFP-reporters with SUN2. e Alternative splicing of sororin intron 1 upon SUN2 knockdown. Experiments were performed in biological triplicates. Significance was determined using a Student’s T-test. * p
    Figure Legend Snippet: SNRPD2, SNRPD3 and NHP2L1 protein interactions. a Characterization of Hs578T GFP BAC-reporter cell lines for splicing factors SNRPD2, SNRPD3 and NHP2L1. Images were captured 72 h after transfection. Scale bar = 30 μm. b Overlap of proteins residing in SNRPD2, SNRPD3 and NHP2L1 complexes, respectively. Splicing factors belonging to the nine candidates selected from the primary screen are labeled in purple. Orange: proteins that were selected for further validation. c Nuclear phenotype 72 h after SUN2 knockdown in Hs578T and MDA-MB-231 cells. Scale bar = 30 μm. d Co-immunoprecipitation of SNRPD2, SNRPD3 and NHP2L1 Hs578T BAC GFP-reporters with SUN2. e Alternative splicing of sororin intron 1 upon SUN2 knockdown. Experiments were performed in biological triplicates. Significance was determined using a Student’s T-test. * p

    Techniques Used: BAC Assay, Transfection, Labeling, Multiple Displacement Amplification, Immunoprecipitation

    Spliceosome RNAi screen identifies novel splicing regulators of tumor cell proliferation . a Overview of steps used to select splicing factors involved in breast cancer cell proliferation. All assays were performed 72 h after transfection. b Correlation between SRB and nuclei count values (both proliferation measurements) in the primary screen for Hs578T (top) and MDA-MB-231 (bottom) cell lines. c Heatmap showing the Z-scores for splicing factor knockdown effects on proliferation in Hs578T and MDA-MB-231 72 h after knockdown. Splicing factors that upon knockdown inhibited proliferation in both assays and cell lines (highlighted clusters) were selected for further validation. d Nuclei counting Z-scores for all splicing factors in the primary screen. Factors selected for validation are highlighted in blue and red, respectively. e Examples of splicing factor knockdowns and nuclei counting. siKinasePool (siKP) was used as a negative control. Scale bar = 500 μm
    Figure Legend Snippet: Spliceosome RNAi screen identifies novel splicing regulators of tumor cell proliferation . a Overview of steps used to select splicing factors involved in breast cancer cell proliferation. All assays were performed 72 h after transfection. b Correlation between SRB and nuclei count values (both proliferation measurements) in the primary screen for Hs578T (top) and MDA-MB-231 (bottom) cell lines. c Heatmap showing the Z-scores for splicing factor knockdown effects on proliferation in Hs578T and MDA-MB-231 72 h after knockdown. Splicing factors that upon knockdown inhibited proliferation in both assays and cell lines (highlighted clusters) were selected for further validation. d Nuclei counting Z-scores for all splicing factors in the primary screen. Factors selected for validation are highlighted in blue and red, respectively. e Examples of splicing factor knockdowns and nuclei counting. siKinasePool (siKP) was used as a negative control. Scale bar = 500 μm

    Techniques Used: Transfection, Sulforhodamine B Assay, Multiple Displacement Amplification, Negative Control

    Effect of candidate splicing factor knockdown on cell death and cell cycle progression . a Results of validation screen; effect of SMARTpool and 4 single siRNAs of selected candidates on proliferation (SRB and nuclei count) and cell death (Annexin V and PI staining) in Hs578T cells 72 h after knockdown. PPIH, SRPK2 and SRRT were splicing factors not affecting proliferation in the primary screen and used as negative control. b Effect of splicing factor knockdown on cell DNA content in Hs578T cells measured by FACS analysis 72 h after knockdown. Mean + stdev of three biological replicates. siSRRT was used as a splicing factor negative control. c Effect of selected splicing factor knockdown on cell cycle arrest measured using the FUCCI cell cycle system in Hs578T cells 72 h after knockdown. Mean + stdev of three biological replicates. siSRRT was used as a splicing factor negative control. d Example of the effect of splicing factor SNRPD3 knockdown on FUCCI cell cycle markers in Hs578T cells. e Effect of splicing factor knockdown on expression levels of cell cycle regulators in Hs578T cells. Representative blots of two biological replicates. f Representative images of propidium iodide (PI) and Hoechst staining 2, 4 and 7 days after knockdown in Hs578T cells. Scale bar = 100 μm. g Percentage of cell death 2, 4 and 7 days after splicing factor knockdown in Hs578T (top) and MDA-MB-231 (bottom) cell lines. Mean + stdev of three biological replicates. h Effect of splicing factor knockdown on caspase activity in Hs578T (top) and MDA-MB-231 (bottom) cell lines 4 days after transfection. Mean + stdev of three biological replicates. Statistical significance was determined using ANOVA correcting for multiple testing. * p
    Figure Legend Snippet: Effect of candidate splicing factor knockdown on cell death and cell cycle progression . a Results of validation screen; effect of SMARTpool and 4 single siRNAs of selected candidates on proliferation (SRB and nuclei count) and cell death (Annexin V and PI staining) in Hs578T cells 72 h after knockdown. PPIH, SRPK2 and SRRT were splicing factors not affecting proliferation in the primary screen and used as negative control. b Effect of splicing factor knockdown on cell DNA content in Hs578T cells measured by FACS analysis 72 h after knockdown. Mean + stdev of three biological replicates. siSRRT was used as a splicing factor negative control. c Effect of selected splicing factor knockdown on cell cycle arrest measured using the FUCCI cell cycle system in Hs578T cells 72 h after knockdown. Mean + stdev of three biological replicates. siSRRT was used as a splicing factor negative control. d Example of the effect of splicing factor SNRPD3 knockdown on FUCCI cell cycle markers in Hs578T cells. e Effect of splicing factor knockdown on expression levels of cell cycle regulators in Hs578T cells. Representative blots of two biological replicates. f Representative images of propidium iodide (PI) and Hoechst staining 2, 4 and 7 days after knockdown in Hs578T cells. Scale bar = 100 μm. g Percentage of cell death 2, 4 and 7 days after splicing factor knockdown in Hs578T (top) and MDA-MB-231 (bottom) cell lines. Mean + stdev of three biological replicates. h Effect of splicing factor knockdown on caspase activity in Hs578T (top) and MDA-MB-231 (bottom) cell lines 4 days after transfection. Mean + stdev of three biological replicates. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Techniques Used: Sulforhodamine B Assay, Staining, Negative Control, FACS, Expressing, Multiple Displacement Amplification, Activity Assay, Transfection

    Candidate splicing factor knockdown results in sororin intron 1 retention leading to reduction of sororin protein levels . a Nuclear phenotype 72 h after candidate splicing factor or sororin knockdown in MDA-MB-231 and Hs578T cells. Scale bar = 50 μm. b Sororin intron 1 (i) and intron 2 (ii) retention 72 h after SNRPD2, SNRPD3 and NHP2L1 knockdown in Hs578T and MDA-MB-231 cells. Mean + stdev of 3 biological replicates. c Sororin intron 1 retention 72 h after knockdown of other splicing factor candidates in Hs578T and MDA-MB-231 cells. Mean + stdev of 3 biological replicates. d Confocal images of MDA-MB-231 cells 24 h after transfection with mScarlet, mScarlet tagged sororin (without introns) or mScarlet tagged sororin with intron 1 retained. mScarlet transfections were combined with GFP vectors to control for transfection efficiency. Scale bar = 100 μm. e Alternative splicing of sororin 24 h after transfection with different mScarlet-Sororin plasmids. Forward primer bound to mScarlet, reversed primer bound to sororin exon 2. f Sororin protein expression 24 h after co-transfection with mScarlet and GFP plasmids. Statistical significance was determined using ANOVA correcting for multiple testing. * p
    Figure Legend Snippet: Candidate splicing factor knockdown results in sororin intron 1 retention leading to reduction of sororin protein levels . a Nuclear phenotype 72 h after candidate splicing factor or sororin knockdown in MDA-MB-231 and Hs578T cells. Scale bar = 50 μm. b Sororin intron 1 (i) and intron 2 (ii) retention 72 h after SNRPD2, SNRPD3 and NHP2L1 knockdown in Hs578T and MDA-MB-231 cells. Mean + stdev of 3 biological replicates. c Sororin intron 1 retention 72 h after knockdown of other splicing factor candidates in Hs578T and MDA-MB-231 cells. Mean + stdev of 3 biological replicates. d Confocal images of MDA-MB-231 cells 24 h after transfection with mScarlet, mScarlet tagged sororin (without introns) or mScarlet tagged sororin with intron 1 retained. mScarlet transfections were combined with GFP vectors to control for transfection efficiency. Scale bar = 100 μm. e Alternative splicing of sororin 24 h after transfection with different mScarlet-Sororin plasmids. Forward primer bound to mScarlet, reversed primer bound to sororin exon 2. f Sororin protein expression 24 h after co-transfection with mScarlet and GFP plasmids. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Techniques Used: Multiple Displacement Amplification, Transfection, Expressing, Cotransfection

    Effect of candidate splicing factor knockdown on sister chromatid cohesion . a Effect of splicing factor knockdown on nuclear phenotype in Hs578T cells 72 h after knockdown. Scale bar = 50 μm. b Percentage of cells displaying an abnormal nuclear phenotype 72 h after knockdown. Mean + stdev of at least 3 positions. c Percentage of p-Histone H3 positive cells upon splicing factor knockdown 72 h after knockdown. Nocodazole treatment was used as a positive control. Mean + stdev of 3 biological replicates. d Nuclear phenotype of p-histone H3 positive cells upon SNRPD2 knockdown in MDA-MB-231. Some cells with irregular nuclear phenotype are p-histone H3 positive (blue squares), while some are negative (white squares). Scale bar = 50 μm. e Overview of the factors involved in sister chromatid cohesion. Adapted from Peters et al, 2012. f Effect of SNRPD2, SNRPD3 and NHP2L1 knockdown on RNA expression levels of sororin, ESPL1, MAU2 and SMC1 in Hs578T 72 h after knockdown. Mean + stdev of 3 biological replicates. g Effect of SNRPD2, SNRPD3 and NHP2L1 knockdown on sororin, ESPL1, MAU2 and SMC1 expression levels 1, 2 and 3 days after knockdown in Hs578T. Mean + stdev of three biological replicates. (H) Protein MAU2, SMC1 and sororin levels 3 days after knockdown of SNRPD2, SNRPD3 and NHP2L1. Statistical significance was determined using ANOVA correcting for multiple testing. * p
    Figure Legend Snippet: Effect of candidate splicing factor knockdown on sister chromatid cohesion . a Effect of splicing factor knockdown on nuclear phenotype in Hs578T cells 72 h after knockdown. Scale bar = 50 μm. b Percentage of cells displaying an abnormal nuclear phenotype 72 h after knockdown. Mean + stdev of at least 3 positions. c Percentage of p-Histone H3 positive cells upon splicing factor knockdown 72 h after knockdown. Nocodazole treatment was used as a positive control. Mean + stdev of 3 biological replicates. d Nuclear phenotype of p-histone H3 positive cells upon SNRPD2 knockdown in MDA-MB-231. Some cells with irregular nuclear phenotype are p-histone H3 positive (blue squares), while some are negative (white squares). Scale bar = 50 μm. e Overview of the factors involved in sister chromatid cohesion. Adapted from Peters et al, 2012. f Effect of SNRPD2, SNRPD3 and NHP2L1 knockdown on RNA expression levels of sororin, ESPL1, MAU2 and SMC1 in Hs578T 72 h after knockdown. Mean + stdev of 3 biological replicates. g Effect of SNRPD2, SNRPD3 and NHP2L1 knockdown on sororin, ESPL1, MAU2 and SMC1 expression levels 1, 2 and 3 days after knockdown in Hs578T. Mean + stdev of three biological replicates. (H) Protein MAU2, SMC1 and sororin levels 3 days after knockdown of SNRPD2, SNRPD3 and NHP2L1. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Techniques Used: Positive Control, Multiple Displacement Amplification, RNA Expression, Expressing

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • hs578t  (ATCC)
    97
    ATCC hs578t
    SNRPD2, SNRPD3 and NHP2L1 protein interactions. a Characterization of <t>Hs578T</t> GFP BAC-reporter cell lines for splicing factors SNRPD2, SNRPD3 and NHP2L1. Images were captured 72 h after transfection. Scale bar = 30 μm. b Overlap of proteins residing in SNRPD2, SNRPD3 and NHP2L1 complexes, respectively. Splicing factors belonging to the nine candidates selected from the primary screen are labeled in purple. Orange: proteins that were selected for further validation. c Nuclear phenotype 72 h after SUN2 knockdown in Hs578T and MDA-MB-231 cells. Scale bar = 30 μm. d Co-immunoprecipitation of SNRPD2, SNRPD3 and NHP2L1 Hs578T BAC GFP-reporters with SUN2. e Alternative splicing of sororin intron 1 upon SUN2 knockdown. Experiments were performed in biological triplicates. Significance was determined using a Student’s T-test. * p
    Hs578t, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hs578t/product/ATCC
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hs578t - by Bioz Stars, 2022-10
    97/100 stars
      Buy from Supplier

    Image Search Results


    SNRPD2, SNRPD3 and NHP2L1 protein interactions. a Characterization of Hs578T GFP BAC-reporter cell lines for splicing factors SNRPD2, SNRPD3 and NHP2L1. Images were captured 72 h after transfection. Scale bar = 30 μm. b Overlap of proteins residing in SNRPD2, SNRPD3 and NHP2L1 complexes, respectively. Splicing factors belonging to the nine candidates selected from the primary screen are labeled in purple. Orange: proteins that were selected for further validation. c Nuclear phenotype 72 h after SUN2 knockdown in Hs578T and MDA-MB-231 cells. Scale bar = 30 μm. d Co-immunoprecipitation of SNRPD2, SNRPD3 and NHP2L1 Hs578T BAC GFP-reporters with SUN2. e Alternative splicing of sororin intron 1 upon SUN2 knockdown. Experiments were performed in biological triplicates. Significance was determined using a Student’s T-test. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention

    doi: 10.1186/s13046-021-01863-4

    Figure Lengend Snippet: SNRPD2, SNRPD3 and NHP2L1 protein interactions. a Characterization of Hs578T GFP BAC-reporter cell lines for splicing factors SNRPD2, SNRPD3 and NHP2L1. Images were captured 72 h after transfection. Scale bar = 30 μm. b Overlap of proteins residing in SNRPD2, SNRPD3 and NHP2L1 complexes, respectively. Splicing factors belonging to the nine candidates selected from the primary screen are labeled in purple. Orange: proteins that were selected for further validation. c Nuclear phenotype 72 h after SUN2 knockdown in Hs578T and MDA-MB-231 cells. Scale bar = 30 μm. d Co-immunoprecipitation of SNRPD2, SNRPD3 and NHP2L1 Hs578T BAC GFP-reporters with SUN2. e Alternative splicing of sororin intron 1 upon SUN2 knockdown. Experiments were performed in biological triplicates. Significance was determined using a Student’s T-test. * p

    Article Snippet: Hs578T (ATCC-HBT-126), MDA-MB-231 (ATCC-HBT-26), HCC1806 (ATCC-CRL-2335), MDA-MB-468 (ATCC-HTB-132), MCF7 (ATCC-HTB-22) and T47D (ATCC-HTB-133) were purchased from ATCC.

    Techniques: BAC Assay, Transfection, Labeling, Multiple Displacement Amplification, Immunoprecipitation

    Spliceosome RNAi screen identifies novel splicing regulators of tumor cell proliferation . a Overview of steps used to select splicing factors involved in breast cancer cell proliferation. All assays were performed 72 h after transfection. b Correlation between SRB and nuclei count values (both proliferation measurements) in the primary screen for Hs578T (top) and MDA-MB-231 (bottom) cell lines. c Heatmap showing the Z-scores for splicing factor knockdown effects on proliferation in Hs578T and MDA-MB-231 72 h after knockdown. Splicing factors that upon knockdown inhibited proliferation in both assays and cell lines (highlighted clusters) were selected for further validation. d Nuclei counting Z-scores for all splicing factors in the primary screen. Factors selected for validation are highlighted in blue and red, respectively. e Examples of splicing factor knockdowns and nuclei counting. siKinasePool (siKP) was used as a negative control. Scale bar = 500 μm

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention

    doi: 10.1186/s13046-021-01863-4

    Figure Lengend Snippet: Spliceosome RNAi screen identifies novel splicing regulators of tumor cell proliferation . a Overview of steps used to select splicing factors involved in breast cancer cell proliferation. All assays were performed 72 h after transfection. b Correlation between SRB and nuclei count values (both proliferation measurements) in the primary screen for Hs578T (top) and MDA-MB-231 (bottom) cell lines. c Heatmap showing the Z-scores for splicing factor knockdown effects on proliferation in Hs578T and MDA-MB-231 72 h after knockdown. Splicing factors that upon knockdown inhibited proliferation in both assays and cell lines (highlighted clusters) were selected for further validation. d Nuclei counting Z-scores for all splicing factors in the primary screen. Factors selected for validation are highlighted in blue and red, respectively. e Examples of splicing factor knockdowns and nuclei counting. siKinasePool (siKP) was used as a negative control. Scale bar = 500 μm

    Article Snippet: Hs578T (ATCC-HBT-126), MDA-MB-231 (ATCC-HBT-26), HCC1806 (ATCC-CRL-2335), MDA-MB-468 (ATCC-HTB-132), MCF7 (ATCC-HTB-22) and T47D (ATCC-HTB-133) were purchased from ATCC.

    Techniques: Transfection, Sulforhodamine B Assay, Multiple Displacement Amplification, Negative Control

    Effect of candidate splicing factor knockdown on cell death and cell cycle progression . a Results of validation screen; effect of SMARTpool and 4 single siRNAs of selected candidates on proliferation (SRB and nuclei count) and cell death (Annexin V and PI staining) in Hs578T cells 72 h after knockdown. PPIH, SRPK2 and SRRT were splicing factors not affecting proliferation in the primary screen and used as negative control. b Effect of splicing factor knockdown on cell DNA content in Hs578T cells measured by FACS analysis 72 h after knockdown. Mean + stdev of three biological replicates. siSRRT was used as a splicing factor negative control. c Effect of selected splicing factor knockdown on cell cycle arrest measured using the FUCCI cell cycle system in Hs578T cells 72 h after knockdown. Mean + stdev of three biological replicates. siSRRT was used as a splicing factor negative control. d Example of the effect of splicing factor SNRPD3 knockdown on FUCCI cell cycle markers in Hs578T cells. e Effect of splicing factor knockdown on expression levels of cell cycle regulators in Hs578T cells. Representative blots of two biological replicates. f Representative images of propidium iodide (PI) and Hoechst staining 2, 4 and 7 days after knockdown in Hs578T cells. Scale bar = 100 μm. g Percentage of cell death 2, 4 and 7 days after splicing factor knockdown in Hs578T (top) and MDA-MB-231 (bottom) cell lines. Mean + stdev of three biological replicates. h Effect of splicing factor knockdown on caspase activity in Hs578T (top) and MDA-MB-231 (bottom) cell lines 4 days after transfection. Mean + stdev of three biological replicates. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention

    doi: 10.1186/s13046-021-01863-4

    Figure Lengend Snippet: Effect of candidate splicing factor knockdown on cell death and cell cycle progression . a Results of validation screen; effect of SMARTpool and 4 single siRNAs of selected candidates on proliferation (SRB and nuclei count) and cell death (Annexin V and PI staining) in Hs578T cells 72 h after knockdown. PPIH, SRPK2 and SRRT were splicing factors not affecting proliferation in the primary screen and used as negative control. b Effect of splicing factor knockdown on cell DNA content in Hs578T cells measured by FACS analysis 72 h after knockdown. Mean + stdev of three biological replicates. siSRRT was used as a splicing factor negative control. c Effect of selected splicing factor knockdown on cell cycle arrest measured using the FUCCI cell cycle system in Hs578T cells 72 h after knockdown. Mean + stdev of three biological replicates. siSRRT was used as a splicing factor negative control. d Example of the effect of splicing factor SNRPD3 knockdown on FUCCI cell cycle markers in Hs578T cells. e Effect of splicing factor knockdown on expression levels of cell cycle regulators in Hs578T cells. Representative blots of two biological replicates. f Representative images of propidium iodide (PI) and Hoechst staining 2, 4 and 7 days after knockdown in Hs578T cells. Scale bar = 100 μm. g Percentage of cell death 2, 4 and 7 days after splicing factor knockdown in Hs578T (top) and MDA-MB-231 (bottom) cell lines. Mean + stdev of three biological replicates. h Effect of splicing factor knockdown on caspase activity in Hs578T (top) and MDA-MB-231 (bottom) cell lines 4 days after transfection. Mean + stdev of three biological replicates. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Article Snippet: Hs578T (ATCC-HBT-126), MDA-MB-231 (ATCC-HBT-26), HCC1806 (ATCC-CRL-2335), MDA-MB-468 (ATCC-HTB-132), MCF7 (ATCC-HTB-22) and T47D (ATCC-HTB-133) were purchased from ATCC.

    Techniques: Sulforhodamine B Assay, Staining, Negative Control, FACS, Expressing, Multiple Displacement Amplification, Activity Assay, Transfection

    Candidate splicing factor knockdown results in sororin intron 1 retention leading to reduction of sororin protein levels . a Nuclear phenotype 72 h after candidate splicing factor or sororin knockdown in MDA-MB-231 and Hs578T cells. Scale bar = 50 μm. b Sororin intron 1 (i) and intron 2 (ii) retention 72 h after SNRPD2, SNRPD3 and NHP2L1 knockdown in Hs578T and MDA-MB-231 cells. Mean + stdev of 3 biological replicates. c Sororin intron 1 retention 72 h after knockdown of other splicing factor candidates in Hs578T and MDA-MB-231 cells. Mean + stdev of 3 biological replicates. d Confocal images of MDA-MB-231 cells 24 h after transfection with mScarlet, mScarlet tagged sororin (without introns) or mScarlet tagged sororin with intron 1 retained. mScarlet transfections were combined with GFP vectors to control for transfection efficiency. Scale bar = 100 μm. e Alternative splicing of sororin 24 h after transfection with different mScarlet-Sororin plasmids. Forward primer bound to mScarlet, reversed primer bound to sororin exon 2. f Sororin protein expression 24 h after co-transfection with mScarlet and GFP plasmids. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention

    doi: 10.1186/s13046-021-01863-4

    Figure Lengend Snippet: Candidate splicing factor knockdown results in sororin intron 1 retention leading to reduction of sororin protein levels . a Nuclear phenotype 72 h after candidate splicing factor or sororin knockdown in MDA-MB-231 and Hs578T cells. Scale bar = 50 μm. b Sororin intron 1 (i) and intron 2 (ii) retention 72 h after SNRPD2, SNRPD3 and NHP2L1 knockdown in Hs578T and MDA-MB-231 cells. Mean + stdev of 3 biological replicates. c Sororin intron 1 retention 72 h after knockdown of other splicing factor candidates in Hs578T and MDA-MB-231 cells. Mean + stdev of 3 biological replicates. d Confocal images of MDA-MB-231 cells 24 h after transfection with mScarlet, mScarlet tagged sororin (without introns) or mScarlet tagged sororin with intron 1 retained. mScarlet transfections were combined with GFP vectors to control for transfection efficiency. Scale bar = 100 μm. e Alternative splicing of sororin 24 h after transfection with different mScarlet-Sororin plasmids. Forward primer bound to mScarlet, reversed primer bound to sororin exon 2. f Sororin protein expression 24 h after co-transfection with mScarlet and GFP plasmids. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Article Snippet: Hs578T (ATCC-HBT-126), MDA-MB-231 (ATCC-HBT-26), HCC1806 (ATCC-CRL-2335), MDA-MB-468 (ATCC-HTB-132), MCF7 (ATCC-HTB-22) and T47D (ATCC-HTB-133) were purchased from ATCC.

    Techniques: Multiple Displacement Amplification, Transfection, Expressing, Cotransfection

    Effect of candidate splicing factor knockdown on sister chromatid cohesion . a Effect of splicing factor knockdown on nuclear phenotype in Hs578T cells 72 h after knockdown. Scale bar = 50 μm. b Percentage of cells displaying an abnormal nuclear phenotype 72 h after knockdown. Mean + stdev of at least 3 positions. c Percentage of p-Histone H3 positive cells upon splicing factor knockdown 72 h after knockdown. Nocodazole treatment was used as a positive control. Mean + stdev of 3 biological replicates. d Nuclear phenotype of p-histone H3 positive cells upon SNRPD2 knockdown in MDA-MB-231. Some cells with irregular nuclear phenotype are p-histone H3 positive (blue squares), while some are negative (white squares). Scale bar = 50 μm. e Overview of the factors involved in sister chromatid cohesion. Adapted from Peters et al, 2012. f Effect of SNRPD2, SNRPD3 and NHP2L1 knockdown on RNA expression levels of sororin, ESPL1, MAU2 and SMC1 in Hs578T 72 h after knockdown. Mean + stdev of 3 biological replicates. g Effect of SNRPD2, SNRPD3 and NHP2L1 knockdown on sororin, ESPL1, MAU2 and SMC1 expression levels 1, 2 and 3 days after knockdown in Hs578T. Mean + stdev of three biological replicates. (H) Protein MAU2, SMC1 and sororin levels 3 days after knockdown of SNRPD2, SNRPD3 and NHP2L1. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Splicing factors control triple-negative breast cancer cell mitosis through SUN2 interaction and sororin intron retention

    doi: 10.1186/s13046-021-01863-4

    Figure Lengend Snippet: Effect of candidate splicing factor knockdown on sister chromatid cohesion . a Effect of splicing factor knockdown on nuclear phenotype in Hs578T cells 72 h after knockdown. Scale bar = 50 μm. b Percentage of cells displaying an abnormal nuclear phenotype 72 h after knockdown. Mean + stdev of at least 3 positions. c Percentage of p-Histone H3 positive cells upon splicing factor knockdown 72 h after knockdown. Nocodazole treatment was used as a positive control. Mean + stdev of 3 biological replicates. d Nuclear phenotype of p-histone H3 positive cells upon SNRPD2 knockdown in MDA-MB-231. Some cells with irregular nuclear phenotype are p-histone H3 positive (blue squares), while some are negative (white squares). Scale bar = 50 μm. e Overview of the factors involved in sister chromatid cohesion. Adapted from Peters et al, 2012. f Effect of SNRPD2, SNRPD3 and NHP2L1 knockdown on RNA expression levels of sororin, ESPL1, MAU2 and SMC1 in Hs578T 72 h after knockdown. Mean + stdev of 3 biological replicates. g Effect of SNRPD2, SNRPD3 and NHP2L1 knockdown on sororin, ESPL1, MAU2 and SMC1 expression levels 1, 2 and 3 days after knockdown in Hs578T. Mean + stdev of three biological replicates. (H) Protein MAU2, SMC1 and sororin levels 3 days after knockdown of SNRPD2, SNRPD3 and NHP2L1. Statistical significance was determined using ANOVA correcting for multiple testing. * p

    Article Snippet: Hs578T (ATCC-HBT-126), MDA-MB-231 (ATCC-HBT-26), HCC1806 (ATCC-CRL-2335), MDA-MB-468 (ATCC-HTB-132), MCF7 (ATCC-HTB-22) and T47D (ATCC-HTB-133) were purchased from ATCC.

    Techniques: Positive Control, Multiple Displacement Amplification, RNA Expression, Expressing