perk af488  (Cell Signaling Technology Inc)


Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc perk af488
    Perk Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/perk af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    perk af488 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    perk af488  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc perk af488
    Perk Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/perk af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    perk af488 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    tcf1 af488  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc tcf1 af488
    Tcf1 Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tcf1 af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tcf1 af488 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    igg isotype control af488 conjugated  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc igg isotype control af488 conjugated
    Igg Isotype Control Af488 Conjugated, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg isotype control af488 conjugated/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    igg isotype control af488 conjugated - by Bioz Stars, 2024-07
    86/100 stars

    Images

    h3k27ac af488 conjugated  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc h3k27ac af488 conjugated
    ( a ) Representative flow cytometry plots showing the population of Rag2 −/− pro-B cells from young and old bone marrow. Numbers indicate percentage of cells in the gates. Gating strategy was shown in Source Data. ( b ) Average total number of bone marrow cells from tibias and femurs of young and old Rag2 −/− mice (n = 8). Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( c ) Number of pro-B cells after CD19 + selection and CD19 + B220 + sorting from young and old Rag2 −/− mice (n = 9). Each point represents an individual mouse. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( d ) An example locus transitioned from compartment B (young) to compartment A (old) with age. Top tracks show Hi-C PC1 of a segment of chromosome 9 that contains Ncam1 in pro-B cells at different ages; the region corresponding to the Ncam1 locus is expanded below. Normalized tracks for <t>H3K27ac</t> and H3K27me3 ChIP-Seq and RNA-Seq across the Ncam1 locus are shown with IGV. Arhgap20 (right) serves as a control gene located in compartment B that is unaffected by age. ( e ) Dot plots for distance between Foxo1 locus to the nuclear periphery (upper panel) or to the nearest γ-satellite signal (bottom panel). Median is shown in red line. N = 100. Unpaired two-sided t test was used, with P -values indicated. ( f ) Hi-C PC1 showing a segment of chromosome3 containing the Foxo1 locus at different ages.
    H3k27ac Af488 Conjugated, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/h3k27ac af488 conjugated/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    h3k27ac af488 conjugated - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Three-dimensional chromatin reorganization regulates B cell development during ageing"

    Article Title: Three-dimensional chromatin reorganization regulates B cell development during ageing

    Journal: Nature Cell Biology

    doi: 10.1038/s41556-024-01424-9

    ( a ) Representative flow cytometry plots showing the population of Rag2 −/− pro-B cells from young and old bone marrow. Numbers indicate percentage of cells in the gates. Gating strategy was shown in Source Data. ( b ) Average total number of bone marrow cells from tibias and femurs of young and old Rag2 −/− mice (n = 8). Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( c ) Number of pro-B cells after CD19 + selection and CD19 + B220 + sorting from young and old Rag2 −/− mice (n = 9). Each point represents an individual mouse. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( d ) An example locus transitioned from compartment B (young) to compartment A (old) with age. Top tracks show Hi-C PC1 of a segment of chromosome 9 that contains Ncam1 in pro-B cells at different ages; the region corresponding to the Ncam1 locus is expanded below. Normalized tracks for H3K27ac and H3K27me3 ChIP-Seq and RNA-Seq across the Ncam1 locus are shown with IGV. Arhgap20 (right) serves as a control gene located in compartment B that is unaffected by age. ( e ) Dot plots for distance between Foxo1 locus to the nuclear periphery (upper panel) or to the nearest γ-satellite signal (bottom panel). Median is shown in red line. N = 100. Unpaired two-sided t test was used, with P -values indicated. ( f ) Hi-C PC1 showing a segment of chromosome3 containing the Foxo1 locus at different ages.
    Figure Legend Snippet: ( a ) Representative flow cytometry plots showing the population of Rag2 −/− pro-B cells from young and old bone marrow. Numbers indicate percentage of cells in the gates. Gating strategy was shown in Source Data. ( b ) Average total number of bone marrow cells from tibias and femurs of young and old Rag2 −/− mice (n = 8). Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( c ) Number of pro-B cells after CD19 + selection and CD19 + B220 + sorting from young and old Rag2 −/− mice (n = 9). Each point represents an individual mouse. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( d ) An example locus transitioned from compartment B (young) to compartment A (old) with age. Top tracks show Hi-C PC1 of a segment of chromosome 9 that contains Ncam1 in pro-B cells at different ages; the region corresponding to the Ncam1 locus is expanded below. Normalized tracks for H3K27ac and H3K27me3 ChIP-Seq and RNA-Seq across the Ncam1 locus are shown with IGV. Arhgap20 (right) serves as a control gene located in compartment B that is unaffected by age. ( e ) Dot plots for distance between Foxo1 locus to the nuclear periphery (upper panel) or to the nearest γ-satellite signal (bottom panel). Median is shown in red line. N = 100. Unpaired two-sided t test was used, with P -values indicated. ( f ) Hi-C PC1 showing a segment of chromosome3 containing the Foxo1 locus at different ages.

    Techniques Used: Flow Cytometry, Selection, Hi-C, ChIP-sequencing, RNA Sequencing Assay

    a , Combined Hi-C contact linear genomic distances density plots from two replicates of young and old pro-B cells. Colour shading refers to compartments (pink), TADs and loops (green), and close interactions (yellow). b , Example of Hi-C contact heatmaps, visualized with Juicebox using Coverage (sqrt) normalization. Difference heatmap (right) shows regions with increased (red) and decreased interactions (blue) in old pro-B cells. c , PC1 of young and old pro-B cells from Hi-C compartment analysis. Numbers of significantly changed euchromatic (A) and heterochromatic (B) bins of 100 kb are indicated in each quadrant. Significantly changed bins were obtained using a two-pass MD method with a one-sided chi-squared test P < 0.01. d , Difference of aggregate H3K27ac and H3K27me3 ( n = 2) signals within significantly switched bins identified in c . e , Differential expression ( n = 4) for genes associated with each category identified in c from RNA-seq analysis. The top and bottom of each box (in d and e ) represents the 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined by two-sided Wilcoxon signed-rank test. f , Chromatin state of Ebf1 locus in young and old pro-B cells. Top tracks show Hi-C compartment PC1, with orange indicating compartment A and blue indicating compartment B. Lower tracks show normalized ChIP–seq and RNA-seq tracks. Clint1 (right) serves as a control gene located in compartment A that is unaffected by age. g . Hi-C heatmaps of the Ebf1 locus. Interactions with greater than twofold change in old pro-B cells are marked by black circles/arcs (reduced) or the blue square/arc (increased). Numbers represent the ratio of Hi-C PETs in old pro-B cells compared with young pro-B cells. h , Nuclear positioning assayed by FISH. Representative nuclei from young and old pro-B cells and non-B cells from Rag2 −/− mice are shown. Probe colours are as indicated. Dashed lines delineate the nuclear periphery. i , Distance between Ebf1 locus and nuclear periphery in pro-B cells and non-B cells ( n = 100) is shown. The red line indicates the median value; P values based on unpaired two-sided t -tests.
    Figure Legend Snippet: a , Combined Hi-C contact linear genomic distances density plots from two replicates of young and old pro-B cells. Colour shading refers to compartments (pink), TADs and loops (green), and close interactions (yellow). b , Example of Hi-C contact heatmaps, visualized with Juicebox using Coverage (sqrt) normalization. Difference heatmap (right) shows regions with increased (red) and decreased interactions (blue) in old pro-B cells. c , PC1 of young and old pro-B cells from Hi-C compartment analysis. Numbers of significantly changed euchromatic (A) and heterochromatic (B) bins of 100 kb are indicated in each quadrant. Significantly changed bins were obtained using a two-pass MD method with a one-sided chi-squared test P < 0.01. d , Difference of aggregate H3K27ac and H3K27me3 ( n = 2) signals within significantly switched bins identified in c . e , Differential expression ( n = 4) for genes associated with each category identified in c from RNA-seq analysis. The top and bottom of each box (in d and e ) represents the 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined by two-sided Wilcoxon signed-rank test. f , Chromatin state of Ebf1 locus in young and old pro-B cells. Top tracks show Hi-C compartment PC1, with orange indicating compartment A and blue indicating compartment B. Lower tracks show normalized ChIP–seq and RNA-seq tracks. Clint1 (right) serves as a control gene located in compartment A that is unaffected by age. g . Hi-C heatmaps of the Ebf1 locus. Interactions with greater than twofold change in old pro-B cells are marked by black circles/arcs (reduced) or the blue square/arc (increased). Numbers represent the ratio of Hi-C PETs in old pro-B cells compared with young pro-B cells. h , Nuclear positioning assayed by FISH. Representative nuclei from young and old pro-B cells and non-B cells from Rag2 −/− mice are shown. Probe colours are as indicated. Dashed lines delineate the nuclear periphery. i , Distance between Ebf1 locus and nuclear periphery in pro-B cells and non-B cells ( n = 100) is shown. The red line indicates the median value; P values based on unpaired two-sided t -tests.

    Techniques Used: Hi-C, Expressing, RNA Sequencing Assay, ChIP-sequencing

    a , The scatter plot on the left displays a quantitative comparison of TAD strength derived from Hi-C data of young and old Rag2 −/− pro-B cells. Each dot in the plot represents one TAD, and TAD strength is represented as the ES, calculated as indicated. TADs showing increased or decreased interactions with age are coloured blue (old specific) or red (young specific), respectively. Dot plot represents reduced TADs in old pro-B cells. TADs encompassing Igh , Ebf1 and Pax5 loci are indicated. Significantly changed TADs were obtained using a two-pass MD method with a one-sided chi-squared test, P < 0.01. b , Difference heatmap showing reduced TAD formation covering the Pax5 gene locus in aged pro-B cells. c – f , Aggregation analysis of H3K27ac ( c ), H3K27me3 ( d ), CTCF ( e ) and Rad21 ( f ), ChIP–seq signal densities at significantly decreased ( n = 240, young-specific group) and increased ( n = 84, old-specific group) TADs identified in a . g , Aggregation analysis of Rad21 ChIP–seq signal densities around peak centre. Rep1 and Rep2 represent two biological experiments. h , Quantitative RT–PCR validation of RNA-seq data ( n = 3). Data are normalized to Copb2 and presented as mean ± standard error of the mean, with each replicate shown as a dot. Unpaired two-sided t -test was used to determine P values. i , Distribution of expression levels for the genes located in significantly changed TADs ( n = 4). The top and bottom of each box represents 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined by two-sided Wilcoxon signed-rank test.
    Figure Legend Snippet: a , The scatter plot on the left displays a quantitative comparison of TAD strength derived from Hi-C data of young and old Rag2 −/− pro-B cells. Each dot in the plot represents one TAD, and TAD strength is represented as the ES, calculated as indicated. TADs showing increased or decreased interactions with age are coloured blue (old specific) or red (young specific), respectively. Dot plot represents reduced TADs in old pro-B cells. TADs encompassing Igh , Ebf1 and Pax5 loci are indicated. Significantly changed TADs were obtained using a two-pass MD method with a one-sided chi-squared test, P < 0.01. b , Difference heatmap showing reduced TAD formation covering the Pax5 gene locus in aged pro-B cells. c – f , Aggregation analysis of H3K27ac ( c ), H3K27me3 ( d ), CTCF ( e ) and Rad21 ( f ), ChIP–seq signal densities at significantly decreased ( n = 240, young-specific group) and increased ( n = 84, old-specific group) TADs identified in a . g , Aggregation analysis of Rad21 ChIP–seq signal densities around peak centre. Rep1 and Rep2 represent two biological experiments. h , Quantitative RT–PCR validation of RNA-seq data ( n = 3). Data are normalized to Copb2 and presented as mean ± standard error of the mean, with each replicate shown as a dot. Unpaired two-sided t -test was used to determine P values. i , Distribution of expression levels for the genes located in significantly changed TADs ( n = 4). The top and bottom of each box represents 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined by two-sided Wilcoxon signed-rank test.

    Techniques Used: Comparison, Derivative Assay, Hi-C, ChIP-sequencing, Quantitative RT-PCR, RNA Sequencing Assay, Expressing

    a , Changes in H3K27ac were identified from ChIP–seq in young and old pro-B cells ( n = 2). Rep1 and Rep2 represent two biological experiments. H3K27ac peaks within 2 kb of the TSS are labelled as promoters (P), others are labelled as enhancers (E). Top, unchanged peaks (shared); middle, peaks that were reduced in old pro-B cells (young selective); and bottom, peaks that were increased in old pro-B cells (old selective). b , Age-associated H3K27ac peaks were annotated to genes; Venn diagram shows overlap between genes containing young- or old-selective H3K27ac peaks. c , Expression levels ( n = 4) of genes containing each H3K27ac peak category. The ‘Other’ category includes genes that are expressed but not annotated to identified H3K27ac peaks. d , Genome browser tracks of genes containing young ( Bcl11a and Foxo1 ) and old-selective ( Cebpa ) H3K27ac peaks. FC, fold change of indicated H3K27ac peak. P values were determined by one-sided Poisson test. e , Aggregation analysis of H3K27ac HiChIP data ( n = 2) with H3K27ac peaks identified in a . The ES for each H3K27ac peak was calculated as shown. The right panels display the average ES for each category of H3K27ac peaks. f , H3K27ac HiChIP interaction ( n = 2) differences for categories identified in part ‘a’ separated by location of H3K27ac at promoter or enhancer. g , Correlation between changes in H3K27ac ChIP–seq signal and H3K27ac HiChIP interaction strength for peaks identified in a . PCC, Pearson correlation coefficient. h , Heatmaps of H3K27ac HiChIP interactions in the Bcl11a locus in young (upper) and old (lower) pro-B cells. The pink arches represent interactions from the Bcl11a promoter. Strength of interactions are denoted by thickness of arches. Top tracks show H3K27ac peaks obtained from ChIP–seq (red) or HiChIP (purple). The top and bottom of each box (in c and f ) represent 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined using two-sided Wilcoxon signed-rank test; asterisks signify P < 1 × 10 −9 .
    Figure Legend Snippet: a , Changes in H3K27ac were identified from ChIP–seq in young and old pro-B cells ( n = 2). Rep1 and Rep2 represent two biological experiments. H3K27ac peaks within 2 kb of the TSS are labelled as promoters (P), others are labelled as enhancers (E). Top, unchanged peaks (shared); middle, peaks that were reduced in old pro-B cells (young selective); and bottom, peaks that were increased in old pro-B cells (old selective). b , Age-associated H3K27ac peaks were annotated to genes; Venn diagram shows overlap between genes containing young- or old-selective H3K27ac peaks. c , Expression levels ( n = 4) of genes containing each H3K27ac peak category. The ‘Other’ category includes genes that are expressed but not annotated to identified H3K27ac peaks. d , Genome browser tracks of genes containing young ( Bcl11a and Foxo1 ) and old-selective ( Cebpa ) H3K27ac peaks. FC, fold change of indicated H3K27ac peak. P values were determined by one-sided Poisson test. e , Aggregation analysis of H3K27ac HiChIP data ( n = 2) with H3K27ac peaks identified in a . The ES for each H3K27ac peak was calculated as shown. The right panels display the average ES for each category of H3K27ac peaks. f , H3K27ac HiChIP interaction ( n = 2) differences for categories identified in part ‘a’ separated by location of H3K27ac at promoter or enhancer. g , Correlation between changes in H3K27ac ChIP–seq signal and H3K27ac HiChIP interaction strength for peaks identified in a . PCC, Pearson correlation coefficient. h , Heatmaps of H3K27ac HiChIP interactions in the Bcl11a locus in young (upper) and old (lower) pro-B cells. The pink arches represent interactions from the Bcl11a promoter. Strength of interactions are denoted by thickness of arches. Top tracks show H3K27ac peaks obtained from ChIP–seq (red) or HiChIP (purple). The top and bottom of each box (in c and f ) represent 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined using two-sided Wilcoxon signed-rank test; asterisks signify P < 1 × 10 −9 .

    Techniques Used: ChIP-sequencing, Expressing, HiChIP

    ( a ) Measurement of H3K27ac levels in Rag2 −/− young (blue) and old (red) pro-B cells via flow cytometry. The IgG isotype controls are shown in dotted lines. The dot plot on the bottom indicates the median fluorescence intensities (MFI) of H3K27ac levels in young and old pro-B cells. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with the P -value indicated. ( b ) Age-selective p300 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. . ( c ) Overlaps of young selective H3K27ac and p300 peaks.( d ) Age-selective Brg1 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. . ( e ) H3K27ac HiChIP heatmaps of the Ebf1 locus in young (upper) and old (lower) pro-B cells. The visualization elements are similar to that shown in Fig. .
    Figure Legend Snippet: ( a ) Measurement of H3K27ac levels in Rag2 −/− young (blue) and old (red) pro-B cells via flow cytometry. The IgG isotype controls are shown in dotted lines. The dot plot on the bottom indicates the median fluorescence intensities (MFI) of H3K27ac levels in young and old pro-B cells. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with the P -value indicated. ( b ) Age-selective p300 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. . ( c ) Overlaps of young selective H3K27ac and p300 peaks.( d ) Age-selective Brg1 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. . ( e ) H3K27ac HiChIP heatmaps of the Ebf1 locus in young (upper) and old (lower) pro-B cells. The visualization elements are similar to that shown in Fig. .

    Techniques Used: Flow Cytometry, Fluorescence, ChIP-sequencing, Binding Assay, HiChIP

    a , Schematic representation of the mouse Igh locus. Regulatory elements are displayed as ovals. Black and red triangles represent CBEs with opposite orientations. b , Difference Hi-C heatmap showing a part of chromosome 12; Igh TAD is boxed. c , Contact frequency heatmaps of the Igh TAD in young and old pro-B cells. A stripe from 3′ CBEs across the Igh locus is indicated by dashed box and zoomed in on the left side of each heatmap. Schematics on top represent the locus to scale. Coloured sections represent 3′ Igh domain (dark blue), proximal V H genes (green), middle V H genes (pink) and distal V H genes (yellow). A difference map for the same region is shown on the right. The red (RI), green (RII) and blue (RIII) bars above the difference map indicate locations of bacterial artificial chromosome (BAC) probes used for FISH assays in f and g . RPKM, reads per kilobase per million mapped reads. d , Age-dependent histone modifications and CTCF and Rad21 binding at the Igh locus. Tracks from replicate young and old pro-B cell samples are shown. e , Quantification of H3K27ac, Rad21, CTCF and H3K27me3 ChIP–seq ( n = 2) signals in the 2 Mb region of the Igh locus that contains V H gene segments. Proximal, middle and distal refer to locations relative to the 3′ end of the locus (C-J-D H ) in part d . ‘Relative signal’ represents read counts per million (CPM) of each region relative to total reads. Bar graph represents the mean of two replicate experiments, with each replicate shown as a dot. f , g , RI, II and III BAC probes (part b) were used in FISH with young and old Rag 2 −/− pro-B cells. Representative nuclei and probe colours are as indicated ( f ). Dot plots of spatial distances between indicated probes ( n = 100) ( g ). Non-B cells represent Rag 2 −/− bone marrow cells depleted of CD19 + pro-B cells. Unpaired two-sided t -test was used for g , with P values indicated.
    Figure Legend Snippet: a , Schematic representation of the mouse Igh locus. Regulatory elements are displayed as ovals. Black and red triangles represent CBEs with opposite orientations. b , Difference Hi-C heatmap showing a part of chromosome 12; Igh TAD is boxed. c , Contact frequency heatmaps of the Igh TAD in young and old pro-B cells. A stripe from 3′ CBEs across the Igh locus is indicated by dashed box and zoomed in on the left side of each heatmap. Schematics on top represent the locus to scale. Coloured sections represent 3′ Igh domain (dark blue), proximal V H genes (green), middle V H genes (pink) and distal V H genes (yellow). A difference map for the same region is shown on the right. The red (RI), green (RII) and blue (RIII) bars above the difference map indicate locations of bacterial artificial chromosome (BAC) probes used for FISH assays in f and g . RPKM, reads per kilobase per million mapped reads. d , Age-dependent histone modifications and CTCF and Rad21 binding at the Igh locus. Tracks from replicate young and old pro-B cell samples are shown. e , Quantification of H3K27ac, Rad21, CTCF and H3K27me3 ChIP–seq ( n = 2) signals in the 2 Mb region of the Igh locus that contains V H gene segments. Proximal, middle and distal refer to locations relative to the 3′ end of the locus (C-J-D H ) in part d . ‘Relative signal’ represents read counts per million (CPM) of each region relative to total reads. Bar graph represents the mean of two replicate experiments, with each replicate shown as a dot. f , g , RI, II and III BAC probes (part b) were used in FISH with young and old Rag 2 −/− pro-B cells. Representative nuclei and probe colours are as indicated ( f ). Dot plots of spatial distances between indicated probes ( n = 100) ( g ). Non-B cells represent Rag 2 −/− bone marrow cells depleted of CD19 + pro-B cells. Unpaired two-sided t -test was used for g , with P values indicated.

    Techniques Used: Hi-C, Binding Assay, ChIP-sequencing

    dapi af488 conjugated phospho histone h3 ser10  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc dapi af488 conjugated phospho histone h3 ser10
    Inhibition of SAC kinase Mps1 bypasses G2/M arrest, but not loss of viability, triggered by FBXO42 inactivation in F42L- S cells. ( A ) Overview of creation of Dox controllable FBXO42 cells. ( B ) Indel analysis of endogenous FBXO42 locus of two GSC-0827 clones. ( C ) Western blot for FBXO42 for clone 15 with either continuous Dox exposure or 4 days after Dox removal. Clone 15 cells harbor a biallelic disruption of endogenous FBXO42 locus and also express a Dox-controlled FBXO42 ORF, which complements the loss of endogenous FBXO42 . ( D ) Photomicrographs taken from time-lapse videos of clone 15 cells grown with and without Dox (times after Dox withdrawal are indicated) (see videos in Supplementary data for full dataset). ( E ) Overview of experiment performed in panel (F). ( F ) DAPI versus phospho-histone H3 <t>(Ser10)</t> flow cytometry profiles for GSC-0827 Dox-inducible FBXO42 clone 15 kept in ±Dox for 4 days and then treated with vehicle or an Mps1 inhibitor (NMS-P715) for 2 h. ( G ) Relative viability for cells that were kept in ±Dox for 4 days and then treated with vehicle or 200 nM Mps1 inhibitor for 22 h ( n = 3; * P val < 0.01, Student’s t -test).
    Dapi Af488 Conjugated Phospho Histone H3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dapi af488 conjugated phospho histone h3 ser10/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dapi af488 conjugated phospho histone h3 ser10 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "FBXO42 activity is required to prevent mitotic arrest, spindle assembly checkpoint activation and lethality in glioblastoma and other cancers"

    Article Title: FBXO42 activity is required to prevent mitotic arrest, spindle assembly checkpoint activation and lethality in glioblastoma and other cancers

    Journal: NAR Cancer

    doi: 10.1093/narcan/zcae021

    Inhibition of SAC kinase Mps1 bypasses G2/M arrest, but not loss of viability, triggered by FBXO42 inactivation in F42L- S cells. ( A ) Overview of creation of Dox controllable FBXO42 cells. ( B ) Indel analysis of endogenous FBXO42 locus of two GSC-0827 clones. ( C ) Western blot for FBXO42 for clone 15 with either continuous Dox exposure or 4 days after Dox removal. Clone 15 cells harbor a biallelic disruption of endogenous FBXO42 locus and also express a Dox-controlled FBXO42 ORF, which complements the loss of endogenous FBXO42 . ( D ) Photomicrographs taken from time-lapse videos of clone 15 cells grown with and without Dox (times after Dox withdrawal are indicated) (see videos in Supplementary data for full dataset). ( E ) Overview of experiment performed in panel (F). ( F ) DAPI versus phospho-histone H3 (Ser10) flow cytometry profiles for GSC-0827 Dox-inducible FBXO42 clone 15 kept in ±Dox for 4 days and then treated with vehicle or an Mps1 inhibitor (NMS-P715) for 2 h. ( G ) Relative viability for cells that were kept in ±Dox for 4 days and then treated with vehicle or 200 nM Mps1 inhibitor for 22 h ( n = 3; * P val < 0.01, Student’s t -test).
    Figure Legend Snippet: Inhibition of SAC kinase Mps1 bypasses G2/M arrest, but not loss of viability, triggered by FBXO42 inactivation in F42L- S cells. ( A ) Overview of creation of Dox controllable FBXO42 cells. ( B ) Indel analysis of endogenous FBXO42 locus of two GSC-0827 clones. ( C ) Western blot for FBXO42 for clone 15 with either continuous Dox exposure or 4 days after Dox removal. Clone 15 cells harbor a biallelic disruption of endogenous FBXO42 locus and also express a Dox-controlled FBXO42 ORF, which complements the loss of endogenous FBXO42 . ( D ) Photomicrographs taken from time-lapse videos of clone 15 cells grown with and without Dox (times after Dox withdrawal are indicated) (see videos in Supplementary data for full dataset). ( E ) Overview of experiment performed in panel (F). ( F ) DAPI versus phospho-histone H3 (Ser10) flow cytometry profiles for GSC-0827 Dox-inducible FBXO42 clone 15 kept in ±Dox for 4 days and then treated with vehicle or an Mps1 inhibitor (NMS-P715) for 2 h. ( G ) Relative viability for cells that were kept in ±Dox for 4 days and then treated with vehicle or 200 nM Mps1 inhibitor for 22 h ( n = 3; * P val < 0.01, Student’s t -test).

    Techniques Used: Inhibition, Clone Assay, Western Blot, Disruption, Flow Cytometry

    tcf1 tcf7 af488  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc tcf1 tcf7 af488
    Tcf1 Tcf7 Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tcf1 tcf7 af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tcf1 tcf7 af488 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    anti rabbit af488  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc anti rabbit af488
    Anti Rabbit Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti rabbit af488 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    af488  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc af488
    (A) CBB staining of Tol 5, its Δ ataA mutant, and its Δ kmtA mutant cell lysates fractionated according to their molecular weight by SDS‒PAGE. The arrowhead shows the position of the AtaA bands. (B) Immunofluorescence microscopy of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody. Fluorescence and bright fields are shown. Scale bar: 2 µm. (C) Fluorescence flow cytometry analysis of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody.
    Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    af488 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "A new target of multiple lysine methylation in bacteria"

    Article Title: A new target of multiple lysine methylation in bacteria

    Journal: bioRxiv

    doi: 10.1101/2024.05.15.594293

    (A) CBB staining of Tol 5, its Δ ataA mutant, and its Δ kmtA mutant cell lysates fractionated according to their molecular weight by SDS‒PAGE. The arrowhead shows the position of the AtaA bands. (B) Immunofluorescence microscopy of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody. Fluorescence and bright fields are shown. Scale bar: 2 µm. (C) Fluorescence flow cytometry analysis of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody.
    Figure Legend Snippet: (A) CBB staining of Tol 5, its Δ ataA mutant, and its Δ kmtA mutant cell lysates fractionated according to their molecular weight by SDS‒PAGE. The arrowhead shows the position of the AtaA bands. (B) Immunofluorescence microscopy of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody. Fluorescence and bright fields are shown. Scale bar: 2 µm. (C) Fluorescence flow cytometry analysis of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody.

    Techniques Used: Staining, Mutagenesis, Molecular Weight, Immunofluorescence, Microscopy, Fluorescence, Flow Cytometry

    anti mouse human tcf1 af488  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc anti mouse human tcf1 af488

    Anti Mouse Human Tcf1 Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mouse human tcf1 af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mouse human tcf1 af488 - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "IL-7-primed bystander CD8 tumor-infiltrating lymphocytes optimize the antitumor efficacy of T cell engager immunotherapy"

    Article Title: IL-7-primed bystander CD8 tumor-infiltrating lymphocytes optimize the antitumor efficacy of T cell engager immunotherapy

    Journal: Cell Reports Medicine

    doi: 10.1016/j.xcrm.2024.101567


    Figure Legend Snippet:

    Techniques Used: Purification, Recombinant, Formulation, Flow Cytometry, Amplification, Software, Staining

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    Cell Signaling Technology Inc perk af488
    Perk Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/perk af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    perk af488 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc tcf1 af488
    Tcf1 Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tcf1 af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tcf1 af488 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc igg isotype control af488 conjugated
    Igg Isotype Control Af488 Conjugated, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg isotype control af488 conjugated/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    igg isotype control af488 conjugated - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc h3k27ac af488 conjugated
    ( a ) Representative flow cytometry plots showing the population of Rag2 −/− pro-B cells from young and old bone marrow. Numbers indicate percentage of cells in the gates. Gating strategy was shown in Source Data. ( b ) Average total number of bone marrow cells from tibias and femurs of young and old Rag2 −/− mice (n = 8). Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( c ) Number of pro-B cells after CD19 + selection and CD19 + B220 + sorting from young and old Rag2 −/− mice (n = 9). Each point represents an individual mouse. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( d ) An example locus transitioned from compartment B (young) to compartment A (old) with age. Top tracks show Hi-C PC1 of a segment of chromosome 9 that contains Ncam1 in pro-B cells at different ages; the region corresponding to the Ncam1 locus is expanded below. Normalized tracks for <t>H3K27ac</t> and H3K27me3 ChIP-Seq and RNA-Seq across the Ncam1 locus are shown with IGV. Arhgap20 (right) serves as a control gene located in compartment B that is unaffected by age. ( e ) Dot plots for distance between Foxo1 locus to the nuclear periphery (upper panel) or to the nearest γ-satellite signal (bottom panel). Median is shown in red line. N = 100. Unpaired two-sided t test was used, with P -values indicated. ( f ) Hi-C PC1 showing a segment of chromosome3 containing the Foxo1 locus at different ages.
    H3k27ac Af488 Conjugated, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/h3k27ac af488 conjugated/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    h3k27ac af488 conjugated - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc dapi af488 conjugated phospho histone h3 ser10
    Inhibition of SAC kinase Mps1 bypasses G2/M arrest, but not loss of viability, triggered by FBXO42 inactivation in F42L- S cells. ( A ) Overview of creation of Dox controllable FBXO42 cells. ( B ) Indel analysis of endogenous FBXO42 locus of two GSC-0827 clones. ( C ) Western blot for FBXO42 for clone 15 with either continuous Dox exposure or 4 days after Dox removal. Clone 15 cells harbor a biallelic disruption of endogenous FBXO42 locus and also express a Dox-controlled FBXO42 ORF, which complements the loss of endogenous FBXO42 . ( D ) Photomicrographs taken from time-lapse videos of clone 15 cells grown with and without Dox (times after Dox withdrawal are indicated) (see videos in Supplementary data for full dataset). ( E ) Overview of experiment performed in panel (F). ( F ) DAPI versus phospho-histone H3 <t>(Ser10)</t> flow cytometry profiles for GSC-0827 Dox-inducible FBXO42 clone 15 kept in ±Dox for 4 days and then treated with vehicle or an Mps1 inhibitor (NMS-P715) for 2 h. ( G ) Relative viability for cells that were kept in ±Dox for 4 days and then treated with vehicle or 200 nM Mps1 inhibitor for 22 h ( n = 3; * P val < 0.01, Student’s t -test).
    Dapi Af488 Conjugated Phospho Histone H3 Ser10, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dapi af488 conjugated phospho histone h3 ser10/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dapi af488 conjugated phospho histone h3 ser10 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc tcf1 tcf7 af488
    Inhibition of SAC kinase Mps1 bypasses G2/M arrest, but not loss of viability, triggered by FBXO42 inactivation in F42L- S cells. ( A ) Overview of creation of Dox controllable FBXO42 cells. ( B ) Indel analysis of endogenous FBXO42 locus of two GSC-0827 clones. ( C ) Western blot for FBXO42 for clone 15 with either continuous Dox exposure or 4 days after Dox removal. Clone 15 cells harbor a biallelic disruption of endogenous FBXO42 locus and also express a Dox-controlled FBXO42 ORF, which complements the loss of endogenous FBXO42 . ( D ) Photomicrographs taken from time-lapse videos of clone 15 cells grown with and without Dox (times after Dox withdrawal are indicated) (see videos in Supplementary data for full dataset). ( E ) Overview of experiment performed in panel (F). ( F ) DAPI versus phospho-histone H3 <t>(Ser10)</t> flow cytometry profiles for GSC-0827 Dox-inducible FBXO42 clone 15 kept in ±Dox for 4 days and then treated with vehicle or an Mps1 inhibitor (NMS-P715) for 2 h. ( G ) Relative viability for cells that were kept in ±Dox for 4 days and then treated with vehicle or 200 nM Mps1 inhibitor for 22 h ( n = 3; * P val < 0.01, Student’s t -test).
    Tcf1 Tcf7 Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tcf1 tcf7 af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tcf1 tcf7 af488 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc anti rabbit af488
    Inhibition of SAC kinase Mps1 bypasses G2/M arrest, but not loss of viability, triggered by FBXO42 inactivation in F42L- S cells. ( A ) Overview of creation of Dox controllable FBXO42 cells. ( B ) Indel analysis of endogenous FBXO42 locus of two GSC-0827 clones. ( C ) Western blot for FBXO42 for clone 15 with either continuous Dox exposure or 4 days after Dox removal. Clone 15 cells harbor a biallelic disruption of endogenous FBXO42 locus and also express a Dox-controlled FBXO42 ORF, which complements the loss of endogenous FBXO42 . ( D ) Photomicrographs taken from time-lapse videos of clone 15 cells grown with and without Dox (times after Dox withdrawal are indicated) (see videos in Supplementary data for full dataset). ( E ) Overview of experiment performed in panel (F). ( F ) DAPI versus phospho-histone H3 <t>(Ser10)</t> flow cytometry profiles for GSC-0827 Dox-inducible FBXO42 clone 15 kept in ±Dox for 4 days and then treated with vehicle or an Mps1 inhibitor (NMS-P715) for 2 h. ( G ) Relative viability for cells that were kept in ±Dox for 4 days and then treated with vehicle or 200 nM Mps1 inhibitor for 22 h ( n = 3; * P val < 0.01, Student’s t -test).
    Anti Rabbit Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti rabbit af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti rabbit af488 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc af488
    (A) CBB staining of Tol 5, its Δ ataA mutant, and its Δ kmtA mutant cell lysates fractionated according to their molecular weight by SDS‒PAGE. The arrowhead shows the position of the AtaA bands. (B) Immunofluorescence microscopy of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody. Fluorescence and bright fields are shown. Scale bar: 2 µm. (C) Fluorescence flow cytometry analysis of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody.
    Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    af488 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc anti mouse human tcf1 af488

    Anti Mouse Human Tcf1 Af488, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mouse human tcf1 af488/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mouse human tcf1 af488 - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    Image Search Results


    ( a ) Representative flow cytometry plots showing the population of Rag2 −/− pro-B cells from young and old bone marrow. Numbers indicate percentage of cells in the gates. Gating strategy was shown in Source Data. ( b ) Average total number of bone marrow cells from tibias and femurs of young and old Rag2 −/− mice (n = 8). Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( c ) Number of pro-B cells after CD19 + selection and CD19 + B220 + sorting from young and old Rag2 −/− mice (n = 9). Each point represents an individual mouse. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( d ) An example locus transitioned from compartment B (young) to compartment A (old) with age. Top tracks show Hi-C PC1 of a segment of chromosome 9 that contains Ncam1 in pro-B cells at different ages; the region corresponding to the Ncam1 locus is expanded below. Normalized tracks for H3K27ac and H3K27me3 ChIP-Seq and RNA-Seq across the Ncam1 locus are shown with IGV. Arhgap20 (right) serves as a control gene located in compartment B that is unaffected by age. ( e ) Dot plots for distance between Foxo1 locus to the nuclear periphery (upper panel) or to the nearest γ-satellite signal (bottom panel). Median is shown in red line. N = 100. Unpaired two-sided t test was used, with P -values indicated. ( f ) Hi-C PC1 showing a segment of chromosome3 containing the Foxo1 locus at different ages.

    Journal: Nature Cell Biology

    Article Title: Three-dimensional chromatin reorganization regulates B cell development during ageing

    doi: 10.1038/s41556-024-01424-9

    Figure Lengend Snippet: ( a ) Representative flow cytometry plots showing the population of Rag2 −/− pro-B cells from young and old bone marrow. Numbers indicate percentage of cells in the gates. Gating strategy was shown in Source Data. ( b ) Average total number of bone marrow cells from tibias and femurs of young and old Rag2 −/− mice (n = 8). Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( c ) Number of pro-B cells after CD19 + selection and CD19 + B220 + sorting from young and old Rag2 −/− mice (n = 9). Each point represents an individual mouse. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with P -values indicated. ( d ) An example locus transitioned from compartment B (young) to compartment A (old) with age. Top tracks show Hi-C PC1 of a segment of chromosome 9 that contains Ncam1 in pro-B cells at different ages; the region corresponding to the Ncam1 locus is expanded below. Normalized tracks for H3K27ac and H3K27me3 ChIP-Seq and RNA-Seq across the Ncam1 locus are shown with IGV. Arhgap20 (right) serves as a control gene located in compartment B that is unaffected by age. ( e ) Dot plots for distance between Foxo1 locus to the nuclear periphery (upper panel) or to the nearest γ-satellite signal (bottom panel). Median is shown in red line. N = 100. Unpaired two-sided t test was used, with P -values indicated. ( f ) Hi-C PC1 showing a segment of chromosome3 containing the Foxo1 locus at different ages.

    Article Snippet: Phycoerythrin (PE) anti-CD19 (BioLegend, 6D5, cat. no. 115508, 1:100), Brilliant Violet 421 (BV421) anti-B220 (BioLegend, RA3-6B2, cat. no. 103240, 1:100), fluorescein isothiocyanate (FITC) anti-IgM (BioLegend, RMM-1, cat. no. 406506; 1:50), PE anti-CD43 (BD Biosciences, S7, cat. no. 553271, 1:100), H3K27ac-AF488 conjugated (Cell Signaling Technology, cat. no. 15485S.

    Techniques: Flow Cytometry, Selection, Hi-C, ChIP-sequencing, RNA Sequencing Assay

    a , Combined Hi-C contact linear genomic distances density plots from two replicates of young and old pro-B cells. Colour shading refers to compartments (pink), TADs and loops (green), and close interactions (yellow). b , Example of Hi-C contact heatmaps, visualized with Juicebox using Coverage (sqrt) normalization. Difference heatmap (right) shows regions with increased (red) and decreased interactions (blue) in old pro-B cells. c , PC1 of young and old pro-B cells from Hi-C compartment analysis. Numbers of significantly changed euchromatic (A) and heterochromatic (B) bins of 100 kb are indicated in each quadrant. Significantly changed bins were obtained using a two-pass MD method with a one-sided chi-squared test P < 0.01. d , Difference of aggregate H3K27ac and H3K27me3 ( n = 2) signals within significantly switched bins identified in c . e , Differential expression ( n = 4) for genes associated with each category identified in c from RNA-seq analysis. The top and bottom of each box (in d and e ) represents the 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined by two-sided Wilcoxon signed-rank test. f , Chromatin state of Ebf1 locus in young and old pro-B cells. Top tracks show Hi-C compartment PC1, with orange indicating compartment A and blue indicating compartment B. Lower tracks show normalized ChIP–seq and RNA-seq tracks. Clint1 (right) serves as a control gene located in compartment A that is unaffected by age. g . Hi-C heatmaps of the Ebf1 locus. Interactions with greater than twofold change in old pro-B cells are marked by black circles/arcs (reduced) or the blue square/arc (increased). Numbers represent the ratio of Hi-C PETs in old pro-B cells compared with young pro-B cells. h , Nuclear positioning assayed by FISH. Representative nuclei from young and old pro-B cells and non-B cells from Rag2 −/− mice are shown. Probe colours are as indicated. Dashed lines delineate the nuclear periphery. i , Distance between Ebf1 locus and nuclear periphery in pro-B cells and non-B cells ( n = 100) is shown. The red line indicates the median value; P values based on unpaired two-sided t -tests.

    Journal: Nature Cell Biology

    Article Title: Three-dimensional chromatin reorganization regulates B cell development during ageing

    doi: 10.1038/s41556-024-01424-9

    Figure Lengend Snippet: a , Combined Hi-C contact linear genomic distances density plots from two replicates of young and old pro-B cells. Colour shading refers to compartments (pink), TADs and loops (green), and close interactions (yellow). b , Example of Hi-C contact heatmaps, visualized with Juicebox using Coverage (sqrt) normalization. Difference heatmap (right) shows regions with increased (red) and decreased interactions (blue) in old pro-B cells. c , PC1 of young and old pro-B cells from Hi-C compartment analysis. Numbers of significantly changed euchromatic (A) and heterochromatic (B) bins of 100 kb are indicated in each quadrant. Significantly changed bins were obtained using a two-pass MD method with a one-sided chi-squared test P < 0.01. d , Difference of aggregate H3K27ac and H3K27me3 ( n = 2) signals within significantly switched bins identified in c . e , Differential expression ( n = 4) for genes associated with each category identified in c from RNA-seq analysis. The top and bottom of each box (in d and e ) represents the 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined by two-sided Wilcoxon signed-rank test. f , Chromatin state of Ebf1 locus in young and old pro-B cells. Top tracks show Hi-C compartment PC1, with orange indicating compartment A and blue indicating compartment B. Lower tracks show normalized ChIP–seq and RNA-seq tracks. Clint1 (right) serves as a control gene located in compartment A that is unaffected by age. g . Hi-C heatmaps of the Ebf1 locus. Interactions with greater than twofold change in old pro-B cells are marked by black circles/arcs (reduced) or the blue square/arc (increased). Numbers represent the ratio of Hi-C PETs in old pro-B cells compared with young pro-B cells. h , Nuclear positioning assayed by FISH. Representative nuclei from young and old pro-B cells and non-B cells from Rag2 −/− mice are shown. Probe colours are as indicated. Dashed lines delineate the nuclear periphery. i , Distance between Ebf1 locus and nuclear periphery in pro-B cells and non-B cells ( n = 100) is shown. The red line indicates the median value; P values based on unpaired two-sided t -tests.

    Article Snippet: Phycoerythrin (PE) anti-CD19 (BioLegend, 6D5, cat. no. 115508, 1:100), Brilliant Violet 421 (BV421) anti-B220 (BioLegend, RA3-6B2, cat. no. 103240, 1:100), fluorescein isothiocyanate (FITC) anti-IgM (BioLegend, RMM-1, cat. no. 406506; 1:50), PE anti-CD43 (BD Biosciences, S7, cat. no. 553271, 1:100), H3K27ac-AF488 conjugated (Cell Signaling Technology, cat. no. 15485S.

    Techniques: Hi-C, Expressing, RNA Sequencing Assay, ChIP-sequencing

    a , The scatter plot on the left displays a quantitative comparison of TAD strength derived from Hi-C data of young and old Rag2 −/− pro-B cells. Each dot in the plot represents one TAD, and TAD strength is represented as the ES, calculated as indicated. TADs showing increased or decreased interactions with age are coloured blue (old specific) or red (young specific), respectively. Dot plot represents reduced TADs in old pro-B cells. TADs encompassing Igh , Ebf1 and Pax5 loci are indicated. Significantly changed TADs were obtained using a two-pass MD method with a one-sided chi-squared test, P < 0.01. b , Difference heatmap showing reduced TAD formation covering the Pax5 gene locus in aged pro-B cells. c – f , Aggregation analysis of H3K27ac ( c ), H3K27me3 ( d ), CTCF ( e ) and Rad21 ( f ), ChIP–seq signal densities at significantly decreased ( n = 240, young-specific group) and increased ( n = 84, old-specific group) TADs identified in a . g , Aggregation analysis of Rad21 ChIP–seq signal densities around peak centre. Rep1 and Rep2 represent two biological experiments. h , Quantitative RT–PCR validation of RNA-seq data ( n = 3). Data are normalized to Copb2 and presented as mean ± standard error of the mean, with each replicate shown as a dot. Unpaired two-sided t -test was used to determine P values. i , Distribution of expression levels for the genes located in significantly changed TADs ( n = 4). The top and bottom of each box represents 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined by two-sided Wilcoxon signed-rank test.

    Journal: Nature Cell Biology

    Article Title: Three-dimensional chromatin reorganization regulates B cell development during ageing

    doi: 10.1038/s41556-024-01424-9

    Figure Lengend Snippet: a , The scatter plot on the left displays a quantitative comparison of TAD strength derived from Hi-C data of young and old Rag2 −/− pro-B cells. Each dot in the plot represents one TAD, and TAD strength is represented as the ES, calculated as indicated. TADs showing increased or decreased interactions with age are coloured blue (old specific) or red (young specific), respectively. Dot plot represents reduced TADs in old pro-B cells. TADs encompassing Igh , Ebf1 and Pax5 loci are indicated. Significantly changed TADs were obtained using a two-pass MD method with a one-sided chi-squared test, P < 0.01. b , Difference heatmap showing reduced TAD formation covering the Pax5 gene locus in aged pro-B cells. c – f , Aggregation analysis of H3K27ac ( c ), H3K27me3 ( d ), CTCF ( e ) and Rad21 ( f ), ChIP–seq signal densities at significantly decreased ( n = 240, young-specific group) and increased ( n = 84, old-specific group) TADs identified in a . g , Aggregation analysis of Rad21 ChIP–seq signal densities around peak centre. Rep1 and Rep2 represent two biological experiments. h , Quantitative RT–PCR validation of RNA-seq data ( n = 3). Data are normalized to Copb2 and presented as mean ± standard error of the mean, with each replicate shown as a dot. Unpaired two-sided t -test was used to determine P values. i , Distribution of expression levels for the genes located in significantly changed TADs ( n = 4). The top and bottom of each box represents 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined by two-sided Wilcoxon signed-rank test.

    Article Snippet: Phycoerythrin (PE) anti-CD19 (BioLegend, 6D5, cat. no. 115508, 1:100), Brilliant Violet 421 (BV421) anti-B220 (BioLegend, RA3-6B2, cat. no. 103240, 1:100), fluorescein isothiocyanate (FITC) anti-IgM (BioLegend, RMM-1, cat. no. 406506; 1:50), PE anti-CD43 (BD Biosciences, S7, cat. no. 553271, 1:100), H3K27ac-AF488 conjugated (Cell Signaling Technology, cat. no. 15485S.

    Techniques: Comparison, Derivative Assay, Hi-C, ChIP-sequencing, Quantitative RT-PCR, RNA Sequencing Assay, Expressing

    a , Changes in H3K27ac were identified from ChIP–seq in young and old pro-B cells ( n = 2). Rep1 and Rep2 represent two biological experiments. H3K27ac peaks within 2 kb of the TSS are labelled as promoters (P), others are labelled as enhancers (E). Top, unchanged peaks (shared); middle, peaks that were reduced in old pro-B cells (young selective); and bottom, peaks that were increased in old pro-B cells (old selective). b , Age-associated H3K27ac peaks were annotated to genes; Venn diagram shows overlap between genes containing young- or old-selective H3K27ac peaks. c , Expression levels ( n = 4) of genes containing each H3K27ac peak category. The ‘Other’ category includes genes that are expressed but not annotated to identified H3K27ac peaks. d , Genome browser tracks of genes containing young ( Bcl11a and Foxo1 ) and old-selective ( Cebpa ) H3K27ac peaks. FC, fold change of indicated H3K27ac peak. P values were determined by one-sided Poisson test. e , Aggregation analysis of H3K27ac HiChIP data ( n = 2) with H3K27ac peaks identified in a . The ES for each H3K27ac peak was calculated as shown. The right panels display the average ES for each category of H3K27ac peaks. f , H3K27ac HiChIP interaction ( n = 2) differences for categories identified in part ‘a’ separated by location of H3K27ac at promoter or enhancer. g , Correlation between changes in H3K27ac ChIP–seq signal and H3K27ac HiChIP interaction strength for peaks identified in a . PCC, Pearson correlation coefficient. h , Heatmaps of H3K27ac HiChIP interactions in the Bcl11a locus in young (upper) and old (lower) pro-B cells. The pink arches represent interactions from the Bcl11a promoter. Strength of interactions are denoted by thickness of arches. Top tracks show H3K27ac peaks obtained from ChIP–seq (red) or HiChIP (purple). The top and bottom of each box (in c and f ) represent 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined using two-sided Wilcoxon signed-rank test; asterisks signify P < 1 × 10 −9 .

    Journal: Nature Cell Biology

    Article Title: Three-dimensional chromatin reorganization regulates B cell development during ageing

    doi: 10.1038/s41556-024-01424-9

    Figure Lengend Snippet: a , Changes in H3K27ac were identified from ChIP–seq in young and old pro-B cells ( n = 2). Rep1 and Rep2 represent two biological experiments. H3K27ac peaks within 2 kb of the TSS are labelled as promoters (P), others are labelled as enhancers (E). Top, unchanged peaks (shared); middle, peaks that were reduced in old pro-B cells (young selective); and bottom, peaks that were increased in old pro-B cells (old selective). b , Age-associated H3K27ac peaks were annotated to genes; Venn diagram shows overlap between genes containing young- or old-selective H3K27ac peaks. c , Expression levels ( n = 4) of genes containing each H3K27ac peak category. The ‘Other’ category includes genes that are expressed but not annotated to identified H3K27ac peaks. d , Genome browser tracks of genes containing young ( Bcl11a and Foxo1 ) and old-selective ( Cebpa ) H3K27ac peaks. FC, fold change of indicated H3K27ac peak. P values were determined by one-sided Poisson test. e , Aggregation analysis of H3K27ac HiChIP data ( n = 2) with H3K27ac peaks identified in a . The ES for each H3K27ac peak was calculated as shown. The right panels display the average ES for each category of H3K27ac peaks. f , H3K27ac HiChIP interaction ( n = 2) differences for categories identified in part ‘a’ separated by location of H3K27ac at promoter or enhancer. g , Correlation between changes in H3K27ac ChIP–seq signal and H3K27ac HiChIP interaction strength for peaks identified in a . PCC, Pearson correlation coefficient. h , Heatmaps of H3K27ac HiChIP interactions in the Bcl11a locus in young (upper) and old (lower) pro-B cells. The pink arches represent interactions from the Bcl11a promoter. Strength of interactions are denoted by thickness of arches. Top tracks show H3K27ac peaks obtained from ChIP–seq (red) or HiChIP (purple). The top and bottom of each box (in c and f ) represent 75th and 25th percentile, respectively, with a line at the median. Whiskers extend by 1.5× the interquartile range. P values were determined using two-sided Wilcoxon signed-rank test; asterisks signify P < 1 × 10 −9 .

    Article Snippet: Phycoerythrin (PE) anti-CD19 (BioLegend, 6D5, cat. no. 115508, 1:100), Brilliant Violet 421 (BV421) anti-B220 (BioLegend, RA3-6B2, cat. no. 103240, 1:100), fluorescein isothiocyanate (FITC) anti-IgM (BioLegend, RMM-1, cat. no. 406506; 1:50), PE anti-CD43 (BD Biosciences, S7, cat. no. 553271, 1:100), H3K27ac-AF488 conjugated (Cell Signaling Technology, cat. no. 15485S.

    Techniques: ChIP-sequencing, Expressing, HiChIP

    ( a ) Measurement of H3K27ac levels in Rag2 −/− young (blue) and old (red) pro-B cells via flow cytometry. The IgG isotype controls are shown in dotted lines. The dot plot on the bottom indicates the median fluorescence intensities (MFI) of H3K27ac levels in young and old pro-B cells. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with the P -value indicated. ( b ) Age-selective p300 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. . ( c ) Overlaps of young selective H3K27ac and p300 peaks.( d ) Age-selective Brg1 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. . ( e ) H3K27ac HiChIP heatmaps of the Ebf1 locus in young (upper) and old (lower) pro-B cells. The visualization elements are similar to that shown in Fig. .

    Journal: Nature Cell Biology

    Article Title: Three-dimensional chromatin reorganization regulates B cell development during ageing

    doi: 10.1038/s41556-024-01424-9

    Figure Lengend Snippet: ( a ) Measurement of H3K27ac levels in Rag2 −/− young (blue) and old (red) pro-B cells via flow cytometry. The IgG isotype controls are shown in dotted lines. The dot plot on the bottom indicates the median fluorescence intensities (MFI) of H3K27ac levels in young and old pro-B cells. Data are presented as mean ± SEM. Unpaired two-sided t test was used, with the P -value indicated. ( b ) Age-selective p300 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. . ( c ) Overlaps of young selective H3K27ac and p300 peaks.( d ) Age-selective Brg1 ChIP-Seq binding sites were identified in young and old pro-B cells. The analysis and cutoffs used were consistent with those in Fig. . ( e ) H3K27ac HiChIP heatmaps of the Ebf1 locus in young (upper) and old (lower) pro-B cells. The visualization elements are similar to that shown in Fig. .

    Article Snippet: Phycoerythrin (PE) anti-CD19 (BioLegend, 6D5, cat. no. 115508, 1:100), Brilliant Violet 421 (BV421) anti-B220 (BioLegend, RA3-6B2, cat. no. 103240, 1:100), fluorescein isothiocyanate (FITC) anti-IgM (BioLegend, RMM-1, cat. no. 406506; 1:50), PE anti-CD43 (BD Biosciences, S7, cat. no. 553271, 1:100), H3K27ac-AF488 conjugated (Cell Signaling Technology, cat. no. 15485S.

    Techniques: Flow Cytometry, Fluorescence, ChIP-sequencing, Binding Assay, HiChIP

    a , Schematic representation of the mouse Igh locus. Regulatory elements are displayed as ovals. Black and red triangles represent CBEs with opposite orientations. b , Difference Hi-C heatmap showing a part of chromosome 12; Igh TAD is boxed. c , Contact frequency heatmaps of the Igh TAD in young and old pro-B cells. A stripe from 3′ CBEs across the Igh locus is indicated by dashed box and zoomed in on the left side of each heatmap. Schematics on top represent the locus to scale. Coloured sections represent 3′ Igh domain (dark blue), proximal V H genes (green), middle V H genes (pink) and distal V H genes (yellow). A difference map for the same region is shown on the right. The red (RI), green (RII) and blue (RIII) bars above the difference map indicate locations of bacterial artificial chromosome (BAC) probes used for FISH assays in f and g . RPKM, reads per kilobase per million mapped reads. d , Age-dependent histone modifications and CTCF and Rad21 binding at the Igh locus. Tracks from replicate young and old pro-B cell samples are shown. e , Quantification of H3K27ac, Rad21, CTCF and H3K27me3 ChIP–seq ( n = 2) signals in the 2 Mb region of the Igh locus that contains V H gene segments. Proximal, middle and distal refer to locations relative to the 3′ end of the locus (C-J-D H ) in part d . ‘Relative signal’ represents read counts per million (CPM) of each region relative to total reads. Bar graph represents the mean of two replicate experiments, with each replicate shown as a dot. f , g , RI, II and III BAC probes (part b) were used in FISH with young and old Rag 2 −/− pro-B cells. Representative nuclei and probe colours are as indicated ( f ). Dot plots of spatial distances between indicated probes ( n = 100) ( g ). Non-B cells represent Rag 2 −/− bone marrow cells depleted of CD19 + pro-B cells. Unpaired two-sided t -test was used for g , with P values indicated.

    Journal: Nature Cell Biology

    Article Title: Three-dimensional chromatin reorganization regulates B cell development during ageing

    doi: 10.1038/s41556-024-01424-9

    Figure Lengend Snippet: a , Schematic representation of the mouse Igh locus. Regulatory elements are displayed as ovals. Black and red triangles represent CBEs with opposite orientations. b , Difference Hi-C heatmap showing a part of chromosome 12; Igh TAD is boxed. c , Contact frequency heatmaps of the Igh TAD in young and old pro-B cells. A stripe from 3′ CBEs across the Igh locus is indicated by dashed box and zoomed in on the left side of each heatmap. Schematics on top represent the locus to scale. Coloured sections represent 3′ Igh domain (dark blue), proximal V H genes (green), middle V H genes (pink) and distal V H genes (yellow). A difference map for the same region is shown on the right. The red (RI), green (RII) and blue (RIII) bars above the difference map indicate locations of bacterial artificial chromosome (BAC) probes used for FISH assays in f and g . RPKM, reads per kilobase per million mapped reads. d , Age-dependent histone modifications and CTCF and Rad21 binding at the Igh locus. Tracks from replicate young and old pro-B cell samples are shown. e , Quantification of H3K27ac, Rad21, CTCF and H3K27me3 ChIP–seq ( n = 2) signals in the 2 Mb region of the Igh locus that contains V H gene segments. Proximal, middle and distal refer to locations relative to the 3′ end of the locus (C-J-D H ) in part d . ‘Relative signal’ represents read counts per million (CPM) of each region relative to total reads. Bar graph represents the mean of two replicate experiments, with each replicate shown as a dot. f , g , RI, II and III BAC probes (part b) were used in FISH with young and old Rag 2 −/− pro-B cells. Representative nuclei and probe colours are as indicated ( f ). Dot plots of spatial distances between indicated probes ( n = 100) ( g ). Non-B cells represent Rag 2 −/− bone marrow cells depleted of CD19 + pro-B cells. Unpaired two-sided t -test was used for g , with P values indicated.

    Article Snippet: Phycoerythrin (PE) anti-CD19 (BioLegend, 6D5, cat. no. 115508, 1:100), Brilliant Violet 421 (BV421) anti-B220 (BioLegend, RA3-6B2, cat. no. 103240, 1:100), fluorescein isothiocyanate (FITC) anti-IgM (BioLegend, RMM-1, cat. no. 406506; 1:50), PE anti-CD43 (BD Biosciences, S7, cat. no. 553271, 1:100), H3K27ac-AF488 conjugated (Cell Signaling Technology, cat. no. 15485S.

    Techniques: Hi-C, Binding Assay, ChIP-sequencing

    Inhibition of SAC kinase Mps1 bypasses G2/M arrest, but not loss of viability, triggered by FBXO42 inactivation in F42L- S cells. ( A ) Overview of creation of Dox controllable FBXO42 cells. ( B ) Indel analysis of endogenous FBXO42 locus of two GSC-0827 clones. ( C ) Western blot for FBXO42 for clone 15 with either continuous Dox exposure or 4 days after Dox removal. Clone 15 cells harbor a biallelic disruption of endogenous FBXO42 locus and also express a Dox-controlled FBXO42 ORF, which complements the loss of endogenous FBXO42 . ( D ) Photomicrographs taken from time-lapse videos of clone 15 cells grown with and without Dox (times after Dox withdrawal are indicated) (see videos in Supplementary data for full dataset). ( E ) Overview of experiment performed in panel (F). ( F ) DAPI versus phospho-histone H3 (Ser10) flow cytometry profiles for GSC-0827 Dox-inducible FBXO42 clone 15 kept in ±Dox for 4 days and then treated with vehicle or an Mps1 inhibitor (NMS-P715) for 2 h. ( G ) Relative viability for cells that were kept in ±Dox for 4 days and then treated with vehicle or 200 nM Mps1 inhibitor for 22 h ( n = 3; * P val < 0.01, Student’s t -test).

    Journal: NAR Cancer

    Article Title: FBXO42 activity is required to prevent mitotic arrest, spindle assembly checkpoint activation and lethality in glioblastoma and other cancers

    doi: 10.1093/narcan/zcae021

    Figure Lengend Snippet: Inhibition of SAC kinase Mps1 bypasses G2/M arrest, but not loss of viability, triggered by FBXO42 inactivation in F42L- S cells. ( A ) Overview of creation of Dox controllable FBXO42 cells. ( B ) Indel analysis of endogenous FBXO42 locus of two GSC-0827 clones. ( C ) Western blot for FBXO42 for clone 15 with either continuous Dox exposure or 4 days after Dox removal. Clone 15 cells harbor a biallelic disruption of endogenous FBXO42 locus and also express a Dox-controlled FBXO42 ORF, which complements the loss of endogenous FBXO42 . ( D ) Photomicrographs taken from time-lapse videos of clone 15 cells grown with and without Dox (times after Dox withdrawal are indicated) (see videos in Supplementary data for full dataset). ( E ) Overview of experiment performed in panel (F). ( F ) DAPI versus phospho-histone H3 (Ser10) flow cytometry profiles for GSC-0827 Dox-inducible FBXO42 clone 15 kept in ±Dox for 4 days and then treated with vehicle or an Mps1 inhibitor (NMS-P715) for 2 h. ( G ) Relative viability for cells that were kept in ±Dox for 4 days and then treated with vehicle or 200 nM Mps1 inhibitor for 22 h ( n = 3; * P val < 0.01, Student’s t -test).

    Article Snippet: Samples were left in ethanol at 4°C overnight and then washed with cold PBS + 2% bovine serum albumin (BSA) and stained for 30 min at 4°C with cold PBS + 2% BSA + 0.1% (v/v) Triton X-100 + 1 μg/ml DAPI + AF488-conjugated phospho-histone H3 (Ser10) antibody (1:100, Cell Signaling Technologies, #3465S).

    Techniques: Inhibition, Clone Assay, Western Blot, Disruption, Flow Cytometry

    (A) CBB staining of Tol 5, its Δ ataA mutant, and its Δ kmtA mutant cell lysates fractionated according to their molecular weight by SDS‒PAGE. The arrowhead shows the position of the AtaA bands. (B) Immunofluorescence microscopy of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody. Fluorescence and bright fields are shown. Scale bar: 2 µm. (C) Fluorescence flow cytometry analysis of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody.

    Journal: bioRxiv

    Article Title: A new target of multiple lysine methylation in bacteria

    doi: 10.1101/2024.05.15.594293

    Figure Lengend Snippet: (A) CBB staining of Tol 5, its Δ ataA mutant, and its Δ kmtA mutant cell lysates fractionated according to their molecular weight by SDS‒PAGE. The arrowhead shows the position of the AtaA bands. (B) Immunofluorescence microscopy of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody. Fluorescence and bright fields are shown. Scale bar: 2 µm. (C) Fluorescence flow cytometry analysis of Tol 5 and its mutants using an anti-AtaA antiserum and an Alexa-Fluor ® 488-conjugated anti-rabbit antibody.

    Article Snippet: The samples were washed twice with PBS-T and incubated with an AF488-conjugated anti-rabbit antibody (Cell Signaling Technology, Inc., Danvers, MA, USA) at a 1:1,000 dilution in PBS-T at room temperature for 30 min.

    Techniques: Staining, Mutagenesis, Molecular Weight, Immunofluorescence, Microscopy, Fluorescence, Flow Cytometry

    Journal: Cell Reports Medicine

    Article Title: IL-7-primed bystander CD8 tumor-infiltrating lymphocytes optimize the antitumor efficacy of T cell engager immunotherapy

    doi: 10.1016/j.xcrm.2024.101567

    Figure Lengend Snippet:

    Article Snippet: Anti-mouse/human TCF1 AF488 (clone: C63D9) , Cell Signaling , Cat# 6444S; RRID: AB_2797627.

    Techniques: Purification, Recombinant, Formulation, Flow Cytometry, Amplification, Software, Staining