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Bethyl phf2
Phf2, supplied by Bethyl, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/phf2/product/Bethyl
Average 94 stars, based on 1 article reviews

phf2 - by Bioz Stars, 2026-04

94/100 stars

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Journal: bioRxiv

Article Title: The AMPKα2/PHF2 axis is critical for turning over lipid droplets during muscle stem cell fate

doi: 10.1101/2025.01.18.630727

Figure Lengend Snippet: CSA distribution of muscle fibers on cryosections of regenerated TA muscles at 11dpi (A) and 28dpi (B). (C) Representative images of immunostainings. (D) Number of myofibers per mm2 at 28dpi. (E) Quantification of the number of PAX7 pos Ki67 neg cells per mm 2 at 28 dpi. (F) Quantification of the number of MYOG pos cells per mm 2 on CTRL SC and PHF2 SCiKO TA cryosections at 11dpi. (G) Percentage of nuclei per myotube, (H) Percentage of PAX7 pos MYOG neg (yellow arrowhead) and (I) PAX7 neg MYOG pos cells (purple arrowhead) in cultures after 48h in LSM. (J) Representative images of immunostainings. (K) Experimental setup. Fusion index (L) and percentage of nuclei per myotube (M) in cultures after 48h in LSM. (N) Experimental setup. Percentage of nuclei per myotube (O) and of PAX7 pos cells (P) in PHF2 SCiKO cells transfected with either MOCK, PHF2 wt or PHF2 H249A . Scale bars, 50 μm. n = 3-6 mice/genotype. n = 3-8 primary cultures/genotype. Values are mean or percentage mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 (Student t -test [D-E-F-H-I-L panels], Sidak’s test after two-way ANOVA [A-B panels], Tukey’s test after one way [G-M-P panels] or two-way ANOVA [O panel]).

Article Snippet: They were transferred in a wet chamber for 1h30 at RT in 1% BSA,50 mM Tris-HCl, pH 8.2 for 1h30 at RT, labelled with gold conjugated Secondary antibody (Goat anti Rabbit 10nm for Myog and Goat anti Rabbit 5nm for Phf2) (Aurion).

Techniques: Muscles, Transfection

(A-B) Percentage of MYOG pos BODIPY pos cells transfected with either MOCK, PHF2 wt or PHF2 S655E and PHF2 S655A . Representative images. (C) Percentage of MYOG pos and (D) percentage of MYOG pos BODIPY pos cells after 48h in LSM. (E) Representative images. (F) Percentage of MYOG pos BODIPY pos cells transfected with either MOCK, PHF2 S655E or PHF2 S655A . Scale bars, 5 μm. n = 3 primary cultures/genotype. Values are mean or percentage mean ± SEM. ***P < 0.001 (Student t -test [C-D panels] and Tukey’s test after one way-ANOVA [B-F panels]).

Journal: bioRxiv

Article Title: The AMPKα2/PHF2 axis is critical for turning over lipid droplets during muscle stem cell fate

doi: 10.1101/2025.01.18.630727

Figure Lengend Snippet: (A-B) Percentage of MYOG pos BODIPY pos cells transfected with either MOCK, PHF2 wt or PHF2 S655E and PHF2 S655A . Representative images. (C) Percentage of MYOG pos and (D) percentage of MYOG pos BODIPY pos cells after 48h in LSM. (E) Representative images. (F) Percentage of MYOG pos BODIPY pos cells transfected with either MOCK, PHF2 S655E or PHF2 S655A . Scale bars, 5 μm. n = 3 primary cultures/genotype. Values are mean or percentage mean ± SEM. ***P < 0.001 (Student t -test [C-D panels] and Tukey’s test after one way-ANOVA [B-F panels]).

Techniques: Transfection

(A) Clustering of EYFP pos -FACS-isolated cells from CTRL SC and PHF2 SCiKO at 11dpi (n=3 mice pooled in 1 sample/genotype). (B) Myog pos myocytes cluster. Plin2, Plin3 (C) and Gdi2 (D) mRNA expression levels. (E) GDI2 protein level in CTRL SC and PHF2 SCiKO mononucleated cells 24h post LSM. (F) Integrative Genomics Viewer (IGV) snapshot depicting PHF2 enrichment (MACS peak call in dark blue) on the Gdi2 promoter region (GSM3462720). PLA analysis (G), electron microscopy analysis of LD-mitochondria contacts (H) and confocal analysis of LD-RAB8a-mitochondria contacts (I) in CTRL SC and PHF2 SCiKO mononucleated cells 24h post LSM. Scale bars, 1, 2 and 5 μm. n = 3 primary cultures/genotype. For (H panel), n=20 cells/genotype/conditions. Values are mean or percentage mean ± SEM. **P < 0.01, ****P < 0.0001 (Student t -test [E panel] and Tukey’s multiple comparison test after one way-ANOVA [G-H-I panels]).

Journal: bioRxiv

Article Title: The AMPKα2/PHF2 axis is critical for turning over lipid droplets during muscle stem cell fate

doi: 10.1101/2025.01.18.630727

Figure Lengend Snippet: (A) Clustering of EYFP pos -FACS-isolated cells from CTRL SC and PHF2 SCiKO at 11dpi (n=3 mice pooled in 1 sample/genotype). (B) Myog pos myocytes cluster. Plin2, Plin3 (C) and Gdi2 (D) mRNA expression levels. (E) GDI2 protein level in CTRL SC and PHF2 SCiKO mononucleated cells 24h post LSM. (F) Integrative Genomics Viewer (IGV) snapshot depicting PHF2 enrichment (MACS peak call in dark blue) on the Gdi2 promoter region (GSM3462720). PLA analysis (G), electron microscopy analysis of LD-mitochondria contacts (H) and confocal analysis of LD-RAB8a-mitochondria contacts (I) in CTRL SC and PHF2 SCiKO mononucleated cells 24h post LSM. Scale bars, 1, 2 and 5 μm. n = 3 primary cultures/genotype. For (H panel), n=20 cells/genotype/conditions. Values are mean or percentage mean ± SEM. **P < 0.01, ****P < 0.0001 (Student t -test [E panel] and Tukey’s multiple comparison test after one way-ANOVA [G-H-I panels]).

Techniques: Isolation, Expressing, Electron Microscopy, Comparison

( A ) Volcano plot of proteins identified by Smc3 ChIP-qMS, normalized to total protein in cohesin immunoprecipitates from WT versus Wapl KO MEFs. The plot shows enriched proteins in the Wapl KO sample on the right and depleted proteins on the left. Biological replicates n = 3. Statistical significance of differentially expressed proteins was determined using limma. Proteins above the purple and red dotted lines represent significance thresholds of p < 0.05 and p < 0.01, respectively. The top enriched protein, Phf2, is highlighted in red. ( B ) Fluorescence microscopy with Phf2 and Scc1 antibodies in WT and Wapl KO MEFs. In the merged panel, Phf2 is shown in green and Scc1 in magenta. Scale bar, 10 μm. ( C ) Live cell microscopy of Phf2-GFP in WT and Wapl KO MEFs. Scale bar, 10 μm. ( D ) Top panel: Immunoblot analysis of whole-cell extracts (WCE) from WT and Wapl KO MEFs. Middle panel: Immunoblot analysis of samples immunoprecipitated (IP) using GFP antibody from WT and Wapl KO MEFs expressing Phf2-GFP. Bottom panel: Silver staining of P samples as described in the middle panel. ( E ) Immunoblot analysis of in vitro binding assay of Phf2 with the cohesin complex. Human recombinant cohesin complexes (Dimer, Trimer, or Tetramer) bound to antibody beads were mixed with purified Phf2. Bound proteins were analyzed using the indicated antibodies. ( F ) Schematic representations of full-length Phf2 and its deletion mutants. PHD, plant homeodomain; JmjC, Jumonji C domain. Δ1:1–450aa; Δ2:451–659aa; Δ3:660–819aa; Δ4:820–1096aa. ( G ) Silver staining of in vitro binding assay of Phf2 deletion mutants with human cohesin. Cohesin tetramer-STAG1 was mixed with purified Phf2 mutants (Δ1, Δ2, Δ3, or Δ4). Flow-through and eluate were analyzed, and the arrowheads marked the positions of the corresponding Phf2 mutants. ( H ) AlphaFold2 model for the interaction between Scc1/Stag1 and Phf2. ( I ) Fluorescence microscopy with Phf2 and Smc3 antibodies in WT and Wapl KO MEF using Phf2 WT, Phf2 Y673A Y675A, Phf2 Y869A Y871A, and Phf2 Δ673–711. In the merged magnified panels, Phf2 is shown in green and Smc3 in magenta. Scale bar, 10 μm. .

Journal: The EMBO Journal

Article Title: Cohesin positions the epigenetic reader Phf2 within the genome

doi: 10.1038/s44318-024-00348-2

Figure Lengend Snippet: ( A ) Volcano plot of proteins identified by Smc3 ChIP-qMS, normalized to total protein in cohesin immunoprecipitates from WT versus Wapl KO MEFs. The plot shows enriched proteins in the Wapl KO sample on the right and depleted proteins on the left. Biological replicates n = 3. Statistical significance of differentially expressed proteins was determined using limma. Proteins above the purple and red dotted lines represent significance thresholds of p < 0.05 and p < 0.01, respectively. The top enriched protein, Phf2, is highlighted in red. ( B ) Fluorescence microscopy with Phf2 and Scc1 antibodies in WT and Wapl KO MEFs. In the merged panel, Phf2 is shown in green and Scc1 in magenta. Scale bar, 10 μm. ( C ) Live cell microscopy of Phf2-GFP in WT and Wapl KO MEFs. Scale bar, 10 μm. ( D ) Top panel: Immunoblot analysis of whole-cell extracts (WCE) from WT and Wapl KO MEFs. Middle panel: Immunoblot analysis of samples immunoprecipitated (IP) using GFP antibody from WT and Wapl KO MEFs expressing Phf2-GFP. Bottom panel: Silver staining of P samples as described in the middle panel. ( E ) Immunoblot analysis of in vitro binding assay of Phf2 with the cohesin complex. Human recombinant cohesin complexes (Dimer, Trimer, or Tetramer) bound to antibody beads were mixed with purified Phf2. Bound proteins were analyzed using the indicated antibodies. ( F ) Schematic representations of full-length Phf2 and its deletion mutants. PHD, plant homeodomain; JmjC, Jumonji C domain. Δ1:1–450aa; Δ2:451–659aa; Δ3:660–819aa; Δ4:820–1096aa. ( G ) Silver staining of in vitro binding assay of Phf2 deletion mutants with human cohesin. Cohesin tetramer-STAG1 was mixed with purified Phf2 mutants (Δ1, Δ2, Δ3, or Δ4). Flow-through and eluate were analyzed, and the arrowheads marked the positions of the corresponding Phf2 mutants. ( H ) AlphaFold2 model for the interaction between Scc1/Stag1 and Phf2. ( I ) Fluorescence microscopy with Phf2 and Smc3 antibodies in WT and Wapl KO MEF using Phf2 WT, Phf2 Y673A Y675A, Phf2 Y869A Y871A, and Phf2 Δ673–711. In the merged magnified panels, Phf2 is shown in green and Smc3 in magenta. Scale bar, 10 μm. .

Article Snippet: The following antibodies were used for immunoblot analysis: rabbit anti-Phf2 (Cell Signaling, 3497S), rabbit anti-Smc3 (Peters laboratory ID A941), mouse anti-Scc1 (Upstate 05-908), rabbit anti-Sa1 (Peters laboratory ID A823), rabbit anti-Sa2 (Peters laboratory ID A824), rabbit anti-Wapl (Peters laboratory ID A960), rabbit anti-CTCF (Peters laboratory ID A992), rabbit anti-MCM3 (Bethyl A300-192A), mouse anti-MCM5 (Santa Cruz sc-136366), mouse anti-Tubulin (Sigma T5168), goat anti-H3 (Santa Cruz sc-8654) and mouse anti-GFP (Roche, 11814460001).

Techniques: Fluorescence, Microscopy, Western Blot, Immunoprecipitation, Expressing, Silver Staining, In Vitro, Binding Assay, Recombinant, Purification

( A ) Binding of Phf2 (in WT and Phf2 KO), H3K4me3, Smc3, and CTCF at a representative locus as determined by ChIP-seq. Small Phf2 peaks colocalizing with Smc3, but not H3K4me3 are indicated with blue arrows. Genes are depicted at the bottom. ( B ) Venn diagram showing the overlap between ChIP-seq peaks of H3K4me3 and Phf2 in WT MEFs. ( C ) Pile-up heat maps and summary plots of ChIP-seq signals obtained for Phf2 (in WT and Phf2 KO MEFs), H3K4me3, Smc3, CTCF, and RNA polymerase II phosphorylated at Ser5 (PolII Ser5-P) shown for the overlap groups indicated in ( B ). The Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. ( D ) Immunoblot analysis of whole-cell extracts from WT and Phf2 KO MEFs using the indicated antibodies. ( E ) Venn diagram showing the overlap between ChIP-seq peaks of Phf2 in WT and Phf2 KO MEFs. ( F ) Venn diagram showing the overlap between ChIP-seq peaks of Smc3 and Phf2 in WT MEFs. ( G ) Pile-up heat maps and summary plots of ChIP-seq signals obtained for Phf2 (in WT and Phf2 KO MEFs), H3K4me3, Smc3, CTCF, and PolII Ser5-P shown for the overlap groups indicated in ( F ). The Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. .

Journal: The EMBO Journal

Article Title: Cohesin positions the epigenetic reader Phf2 within the genome

doi: 10.1038/s44318-024-00348-2

Figure Lengend Snippet: ( A ) Binding of Phf2 (in WT and Phf2 KO), H3K4me3, Smc3, and CTCF at a representative locus as determined by ChIP-seq. Small Phf2 peaks colocalizing with Smc3, but not H3K4me3 are indicated with blue arrows. Genes are depicted at the bottom. ( B ) Venn diagram showing the overlap between ChIP-seq peaks of H3K4me3 and Phf2 in WT MEFs. ( C ) Pile-up heat maps and summary plots of ChIP-seq signals obtained for Phf2 (in WT and Phf2 KO MEFs), H3K4me3, Smc3, CTCF, and RNA polymerase II phosphorylated at Ser5 (PolII Ser5-P) shown for the overlap groups indicated in ( B ). The Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. ( D ) Immunoblot analysis of whole-cell extracts from WT and Phf2 KO MEFs using the indicated antibodies. ( E ) Venn diagram showing the overlap between ChIP-seq peaks of Phf2 in WT and Phf2 KO MEFs. ( F ) Venn diagram showing the overlap between ChIP-seq peaks of Smc3 and Phf2 in WT MEFs. ( G ) Pile-up heat maps and summary plots of ChIP-seq signals obtained for Phf2 (in WT and Phf2 KO MEFs), H3K4me3, Smc3, CTCF, and PolII Ser5-P shown for the overlap groups indicated in ( F ). The Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. .

Techniques: Binding Assay, ChIP-sequencing, Western Blot

( A ) Immunoblot analysis of Phf2 in Phf2 KO MEFs. MEFs with floxed alleles of Phf2 with or without ERCre ( Phf2 F/F, ERCre/+ or Phf2 F/F, no ERCre) were treated with 4-OHT for the indicated days and whole-cell extracts were analyzed using the indicated antibodies. ( B ) Venn diagram illustrating the overlap between ChIP-seq peaks of CTCF and Phf2 in WT MEFs. ( C ) Pile-up heat maps and summary plots of ChIP-seq signals for Phf2 (in WT and Phf2 KO), H3K4me3, Smc3, CTCF, and PolII Ser5-P at overlap groups indicated in ( B ). The Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. ( D ) Summary plots of ChIP-seq signals obtained for Phf2 and Smc3 at oriented CTCF sites. The binding at the CTCF sequence on the + and the – strand was assessed in WT (left) and Wapl KO MEFs (right). .

Journal: The EMBO Journal

Article Title: Cohesin positions the epigenetic reader Phf2 within the genome

doi: 10.1038/s44318-024-00348-2

Figure Lengend Snippet: ( A ) Immunoblot analysis of Phf2 in Phf2 KO MEFs. MEFs with floxed alleles of Phf2 with or without ERCre ( Phf2 F/F, ERCre/+ or Phf2 F/F, no ERCre) were treated with 4-OHT for the indicated days and whole-cell extracts were analyzed using the indicated antibodies. ( B ) Venn diagram illustrating the overlap between ChIP-seq peaks of CTCF and Phf2 in WT MEFs. ( C ) Pile-up heat maps and summary plots of ChIP-seq signals for Phf2 (in WT and Phf2 KO), H3K4me3, Smc3, CTCF, and PolII Ser5-P at overlap groups indicated in ( B ). The Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. ( D ) Summary plots of ChIP-seq signals obtained for Phf2 and Smc3 at oriented CTCF sites. The binding at the CTCF sequence on the + and the – strand was assessed in WT (left) and Wapl KO MEFs (right). .

Techniques: Western Blot, ChIP-sequencing, Binding Assay, Sequencing

$( A ) Immunoblot analysis of soluble (Sol) and chromatin-bound (Chr) fractions from WT and Smc3 KO MEFs using the indicated antibodies. ( B ) Binding of Phf2 (in WT and Smc3 KO MEFs), H3K4me3, Smc3, and CTCF at a representative locus, as determined by ChIP-seq. Small Phf2 peaks detectable in WT, but not in Smc3 KO MEFs are indicated with blue arrows. ( C ) Venn diagram showing the overlap between ChIP-seq peaks of Phf2 in WT and Smc3 KO MEFs. ( D ) Pile-up heat maps and summary plots of ChIP-seq signals obtained for Phf2 (in WT and Smc3 KO MEFs), H3K4me3, Smc3, CTCF, and PolII Ser5-P at the overlap groups indicated in ( C ). The Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. Phf2 peaks present only in WT MEFs were reduced by 64% upon Smc3 depletion, whereas Phf2 peaks that were present in WT and Smc3 KO MEFs were reduced by only 13%. .$

Journal: The EMBO Journal

Article Title: Cohesin positions the epigenetic reader Phf2 within the genome

doi: 10.1038/s44318-024-00348-2

Figure Lengend Snippet: ( A ) Immunoblot analysis of soluble (Sol) and chromatin-bound (Chr) fractions from WT and Smc3 KO MEFs using the indicated antibodies. ( B ) Binding of Phf2 (in WT and Smc3 KO MEFs), H3K4me3, Smc3, and CTCF at a representative locus, as determined by ChIP-seq. Small Phf2 peaks detectable in WT, but not in Smc3 KO MEFs are indicated with blue arrows. ( C ) Venn diagram showing the overlap between ChIP-seq peaks of Phf2 in WT and Smc3 KO MEFs. ( D ) Pile-up heat maps and summary plots of ChIP-seq signals obtained for Phf2 (in WT and Smc3 KO MEFs), H3K4me3, Smc3, CTCF, and PolII Ser5-P at the overlap groups indicated in ( C ). The Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. Phf2 peaks present only in WT MEFs were reduced by 64% upon Smc3 depletion, whereas Phf2 peaks that were present in WT and Smc3 KO MEFs were reduced by only 13%. .

Techniques: Western Blot, Binding Assay, ChIP-sequencing

$( A ) Immunoblot analysis of whole-cell extracts from WT and Smc3 KO MEFs using the indicated antibodies. ( B ) Immunoblot analysis of whole-cell extract (WCE), soluble (Sol), and chromatin-bound (Chr) fractions from WT and Smc3 KO MEFs using the indicated antibodies. Two protein dilutions were assessed. Please note that a portion of this blot was used in Fig. . ( C ) Venn diagram illustrating the overlap between ChIP-seq peaks of Phf2 in WT and Smc3 KO MEFs. ( D ) Pile-up heat maps and summary plots of ChIP-seq signals for Phf2 (in WT and Smc3 KO), H3K4me3, Smc3, CTCF, PolII Ser5-P, and Scc1 (in WT and Smc3 KO; data from Busslinger et al, ) at overlap groups indicated in ( C ). Phf2 peaks present only in WT MEFs were reduced by 64% in read numbers upon Smc3 depletion, whereas Phf2 peaks that were present in WT and Smc3 KO MEFs were reduced by only 13%. Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. The panels from Fig. are shown here again to facilitate a direct side-by-side comparison with the extended panels presented in this figure.$

Journal: The EMBO Journal

Article Title: Cohesin positions the epigenetic reader Phf2 within the genome

doi: 10.1038/s44318-024-00348-2

Figure Lengend Snippet: ( A ) Immunoblot analysis of whole-cell extracts from WT and Smc3 KO MEFs using the indicated antibodies. ( B ) Immunoblot analysis of whole-cell extract (WCE), soluble (Sol), and chromatin-bound (Chr) fractions from WT and Smc3 KO MEFs using the indicated antibodies. Two protein dilutions were assessed. Please note that a portion of this blot was used in Fig. . ( C ) Venn diagram illustrating the overlap between ChIP-seq peaks of Phf2 in WT and Smc3 KO MEFs. ( D ) Pile-up heat maps and summary plots of ChIP-seq signals for Phf2 (in WT and Smc3 KO), H3K4me3, Smc3, CTCF, PolII Ser5-P, and Scc1 (in WT and Smc3 KO; data from Busslinger et al, ) at overlap groups indicated in ( C ). Phf2 peaks present only in WT MEFs were reduced by 64% in read numbers upon Smc3 depletion, whereas Phf2 peaks that were present in WT and Smc3 KO MEFs were reduced by only 13%. Zoom-in panels show the indicated sub-groups of ChIP-seq signals at different color scales. The panels from Fig. are shown here again to facilitate a direct side-by-side comparison with the extended panels presented in this figure.

Techniques: Western Blot, ChIP-sequencing, Comparison

$( A ) Binding of H3K4me3, Phf2 (in WT and Phf2 KO MEFs), Smc3 (in WT and Phf2 KO MEFs), and CTCF at two representative loci, as determined by ChIP-seq. Smc3 peaks detected only in WT, but not in Phf2 KO MEFs are indicated with red arrows. ( B ) Venn diagram showing the overlap between ChIP-seq peaks obtained for Smc3 in WT and Phf2 KO MEFs, with Phf2 peaks in WT MEFs. ( C ) Pile-up summary plots (top) and heat maps (bottom) of ChIP-seq signals obtained for Phf2 in WT MEFs, Smc3 (in WT and Phf2 KO MEFs), and CTCF at the overlap groups indicated in ( B ). Numbers indicate the reduction of ChIP-seq signal of Smc3 peaks in Phf2 KO MEFs. ( D ) Immunoblot analysis of whole-cell extract (WCE), soluble (Sol), and chromatin-bound (Chr) fractions from WT and Phf2 KO MEFs using the indicated antibodies. ( E ) Summary plots of ChIP-seq signals for H3K4me3, Phf2, and CTCF at Smc3 peaks showing a reduction of more than or less than 30% upon Phf2 depletion. ( F ) Summary plots of ChIP-seq signals obtained for H3K4me3, Phf2, Smc3, and CTCF at active TSSs that contain CTCF or not. The purple dotted line indicates the apex of the cumulative CTCF signal, while the blue dotted line indicates the peak of the Phf2 signal. .$

Journal: The EMBO Journal

Article Title: Cohesin positions the epigenetic reader Phf2 within the genome

doi: 10.1038/s44318-024-00348-2

Figure Lengend Snippet: ( A ) Binding of H3K4me3, Phf2 (in WT and Phf2 KO MEFs), Smc3 (in WT and Phf2 KO MEFs), and CTCF at two representative loci, as determined by ChIP-seq. Smc3 peaks detected only in WT, but not in Phf2 KO MEFs are indicated with red arrows. ( B ) Venn diagram showing the overlap between ChIP-seq peaks obtained for Smc3 in WT and Phf2 KO MEFs, with Phf2 peaks in WT MEFs. ( C ) Pile-up summary plots (top) and heat maps (bottom) of ChIP-seq signals obtained for Phf2 in WT MEFs, Smc3 (in WT and Phf2 KO MEFs), and CTCF at the overlap groups indicated in ( B ). Numbers indicate the reduction of ChIP-seq signal of Smc3 peaks in Phf2 KO MEFs. ( D ) Immunoblot analysis of whole-cell extract (WCE), soluble (Sol), and chromatin-bound (Chr) fractions from WT and Phf2 KO MEFs using the indicated antibodies. ( E ) Summary plots of ChIP-seq signals for H3K4me3, Phf2, and CTCF at Smc3 peaks showing a reduction of more than or less than 30% upon Phf2 depletion. ( F ) Summary plots of ChIP-seq signals obtained for H3K4me3, Phf2, Smc3, and CTCF at active TSSs that contain CTCF or not. The purple dotted line indicates the apex of the cumulative CTCF signal, while the blue dotted line indicates the peak of the Phf2 signal. .

Techniques: Binding Assay, ChIP-sequencing, Western Blot

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