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rabbit polyclonal anti wdr5 (Proteintech)

Name: rabbit polyclonal anti wdr5
Brand: Proteintech
Rating: 94 (22 reviews)

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Proteintech rabbit polyclonal anti wdr5
Rabbit Polyclonal Anti Wdr5, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit polyclonal anti wdr5/product/Proteintech
Average 94 stars, based on 22 article reviews

rabbit polyclonal anti wdr5 - by Bioz Stars, 2026-02

94/100 stars

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( A ) MYC transcript levels in control and MEN1-KO cells as determined by RT-qPCR. ns p > 0.05, * p ≤ 0.05. ( B ) MYC transcript counts from RNA-seq of A673 control and MEN1-KO tumors. ( C ) Western blot of MYC and Vinculin levels in A673 and TC32 control and MEN1-KO cells ( D ) Western blot of MYC and Vinculin in tumors derived from A673 control and MEN1-KO cells. ( E ) Co-immunoprecipitations with an anti-Menin antibody were performed on A673, CHLA10 and U2OS nuclear extracts and immunoblotted for Menin, MYC, MLL2 and WDR5. ( F ) Co-immunoprecipitations with an anti-MYC antibody were performed on A673 and CHLA10 nuclear extracts and immunoblotted for MYC, Menin, MAX, MLL2 and WDR5.

Journal: bioRxiv

Article Title: Menin inhibition impairs metastatic colonization of Ewing sarcoma

doi: 10.1101/2025.11.10.687648

Figure Lengend Snippet: ( A ) MYC transcript levels in control and MEN1-KO cells as determined by RT-qPCR. ns p > 0.05, * p ≤ 0.05. ( B ) MYC transcript counts from RNA-seq of A673 control and MEN1-KO tumors. ( C ) Western blot of MYC and Vinculin levels in A673 and TC32 control and MEN1-KO cells ( D ) Western blot of MYC and Vinculin in tumors derived from A673 control and MEN1-KO cells. ( E ) Co-immunoprecipitations with an anti-Menin antibody were performed on A673, CHLA10 and U2OS nuclear extracts and immunoblotted for Menin, MYC, MLL2 and WDR5. ( F ) Co-immunoprecipitations with an anti-MYC antibody were performed on A673 and CHLA10 nuclear extracts and immunoblotted for MYC, Menin, MAX, MLL2 and WDR5.

Article Snippet: Membranes were blocked with Intercept (TBS) Blocking Buffer (LICORbio 927-60001) and probed for the primary antibodies GAPDH Rabbit mAb (Cell Signaling 2118), MAX Rabbit pAb (Cell Signaling 4739), Menin Goat pAb (Bethyl A300-106A), MLL1-C (C-terminal) Rabbit pAb (Bethyl A300-374A), MLL2-C (C-terminal) Rabbit mAb (Cell Signaling 63735), MYC Rabbit mAb (Cell Signaling 18583), Vinculin Rabbit mAb (Cell Signaling 13901) or WDR5 Rabbit pAb (Bethyl A302-430A) followed by the secondary antibody Goat anti-Rabbit 800CW (Licor 926-32211), Donkey anti-Rabbit 800CW (Licor 926-32213) or Donkey anti-Goat 680RD (Licor 926-68074).

Techniques: Control, Quantitative RT-PCR, RNA Sequencing, Western Blot, Derivative Assay

( A ) Incucyte proliferation assay plotting percent confluence for A673 and TC32 cells treated with 0.1% DMSO or 10 µM VTP50469 starting at time 0. Representative of n=3-4). ( B and C ) Invasion of cells from a sphere of A673 cells embedded in rat tail collagen and treated with 0.1% DMSO or 10 µM VTP50469 for 5 days. ( B ) Representative phase and phalloidin (red)/DAPI (blue) stained images are shown (scale bars=100 µm). ( C ) Plot of circularity quantified from replicate spheroids treated as in ( B ). ( D ) A673 and TC32 cells were treated with 0.1% DMSO or 10 µM VTP50469 for 72 hours and genes that were significantly downregulated in VTP50469-treated cells (padj<0.05) were overlapped with genes downregulated in MEN1-KO cells. The top 5 enriched Hallmark pathways for the overlapping downregulated genes in each cell line are shown along with the overlap/total genes for each pathway. The green arrows indicate pathways that were reactivated in A673 MEN1-KO tumors (see ). ( E ) MYC transcript levels in A673, CHLA10 and TC32 cells treated with 0.1% DMSO or 10 µM VTP50469 for 72 hours as determined by RT-qPCR (ns p > 0.05). ( F ) Western of MYC and Vinculin in cells treated with 0.1% DMSO or 10 µM VTP50469 for 72 hours. ( G ) Co-immunoprecipitations with an anti-Menin antibody were performed on nuclear extracts from A673 and CHLA10 cells treated with 0.1% DMSO or 10 µM VTP50469 for 72 hours and immunoblotted for Menin, MYC, MLL2 and WDR5.

Journal: bioRxiv

Article Title: Menin inhibition impairs metastatic colonization of Ewing sarcoma

doi: 10.1101/2025.11.10.687648

Figure Lengend Snippet: ( A ) Incucyte proliferation assay plotting percent confluence for A673 and TC32 cells treated with 0.1% DMSO or 10 µM VTP50469 starting at time 0. Representative of n=3-4). ( B and C ) Invasion of cells from a sphere of A673 cells embedded in rat tail collagen and treated with 0.1% DMSO or 10 µM VTP50469 for 5 days. ( B ) Representative phase and phalloidin (red)/DAPI (blue) stained images are shown (scale bars=100 µm). ( C ) Plot of circularity quantified from replicate spheroids treated as in ( B ). ( D ) A673 and TC32 cells were treated with 0.1% DMSO or 10 µM VTP50469 for 72 hours and genes that were significantly downregulated in VTP50469-treated cells (padj<0.05) were overlapped with genes downregulated in MEN1-KO cells. The top 5 enriched Hallmark pathways for the overlapping downregulated genes in each cell line are shown along with the overlap/total genes for each pathway. The green arrows indicate pathways that were reactivated in A673 MEN1-KO tumors (see ). ( E ) MYC transcript levels in A673, CHLA10 and TC32 cells treated with 0.1% DMSO or 10 µM VTP50469 for 72 hours as determined by RT-qPCR (ns p > 0.05). ( F ) Western of MYC and Vinculin in cells treated with 0.1% DMSO or 10 µM VTP50469 for 72 hours. ( G ) Co-immunoprecipitations with an anti-Menin antibody were performed on nuclear extracts from A673 and CHLA10 cells treated with 0.1% DMSO or 10 µM VTP50469 for 72 hours and immunoblotted for Menin, MYC, MLL2 and WDR5.

Techniques: Proliferation Assay, Staining, Quantitative RT-PCR, Western Blot

( A and B ) Images of colony formation using the indicated human SS cells (HSSY II, SYO-1, Yamato-SS, and MoJo), stably transduced with either a control shRNA or the ones targeting WDR5 (shWDR5-1 or shWDR5-2) (A), or the parental SS cells grown in the presence of the indicated concentration of DMSO, OICR-9429, MS67, MS67N1, or MS67N2 (B). All experiments were repeated at least twice, with representative results shown here. ( C and D ) WB of WDR5, RBPP5, c-MYC, and tubulin in HSSY II (left) and SYO-1 cells (right) treated with the indicated concentration of DMSO, MS67, MS67N1, or MS67N2 for 48 hours (h) (C), or with 2.5 μM of MS67 for the indicated duration (D). All experiments were repeated at least twice, with representative results shown here. ( E ) Plots of growth inhibition using the indicated SS cells, treated with a range of concentration ( x axis) of either OICR-9429 (top), MS67 (second row), MS67N1 (third row), or MS67N2 (bottom) for 2, 4, 6, or 8 days. Y axis, presented in the means ± SEM of replicated data, shows the relative growth after normalization of the cell number to the DMSO-treated controls ( n = 3 independent experiments).

Journal: Science Advances

Article Title: Pharmacologic degradation of WDR5 suppresses oncogenic activities of SS18::SSX and provides a therapeutic of synovial sarcoma

doi: 10.1126/sciadv.ads7876

Figure Lengend Snippet: ( A and B ) Images of colony formation using the indicated human SS cells (HSSY II, SYO-1, Yamato-SS, and MoJo), stably transduced with either a control shRNA or the ones targeting WDR5 (shWDR5-1 or shWDR5-2) (A), or the parental SS cells grown in the presence of the indicated concentration of DMSO, OICR-9429, MS67, MS67N1, or MS67N2 (B). All experiments were repeated at least twice, with representative results shown here. ( C and D ) WB of WDR5, RBPP5, c-MYC, and tubulin in HSSY II (left) and SYO-1 cells (right) treated with the indicated concentration of DMSO, MS67, MS67N1, or MS67N2 for 48 hours (h) (C), or with 2.5 μM of MS67 for the indicated duration (D). All experiments were repeated at least twice, with representative results shown here. ( E ) Plots of growth inhibition using the indicated SS cells, treated with a range of concentration ( x axis) of either OICR-9429 (top), MS67 (second row), MS67N1 (third row), or MS67N2 (bottom) for 2, 4, 6, or 8 days. Y axis, presented in the means ± SEM of replicated data, shows the relative growth after normalization of the cell number to the DMSO-treated controls ( n = 3 independent experiments).

Article Snippet: The primary antibodies used in the study (all diluted at 1:1000) include those against WDR5 (Cell Signaling Technology, 13105), WDR5 (Santa Cruz Biotechnology, sc-393080), SS18::SSX (Cell Signaling Technology, 72364), SS18 (Cell Signaling Technology, 21792), SMARCC1/BAF155 (Cell Signaling Technology, 11956), SMARCA4/BRG1 (Abcam, ab110641), KMT2A/MLL1 (Cell Signaling Technology, 14197), RBBP5 (Cell Signaling Technology, 13171), H3K4me1 (Abcam, ab8895), H3K4me2 (Cell Signaling Technology, 9725), H3K4me3 (Cell Signaling Technology, 9751), general H3 (Abcam, ab1791), cMYC (Abcam, ab32072), P53 (Cell Signaling Technology, 2524), P21 (Cell Signaling Technology, 2947), cleaved caspase 3 (Cell Signaling Technology, 9661), cleaved caspase 7 (Cell Signaling Technology, 8438), glyceraldehyde-3-phosphate dehydrogenase (Cell Signaling Technology, 2118), and tubulin (Cell Signaling Technology, 2146).

Techniques: Stable Transfection, Transduction, Control, shRNA, Concentration Assay, Inhibition

( A ) Representative co-IF images of SS18::SSX, WDR5 and DNA [probed with 4′,6-diamidino-2-phenylindole (DAPI)] in HSSY II cells, treated with 2.5 μM of DMSO (top) or MS67 (bottom) for 48 hours. Scale bar, 5 μm. The experiment was repeated at least twice, with representative results shown here. ( B and C ) co-IP to detect interaction between WDR and the SWI/SNF complex components in HSSY II (B) or U2OS cells (C) using either nonspecific IgG (lane 2), anti-SS18::SSX [lane 3 in (B)], or anti-WDR5 antibody [lane 4 in (B) and lane 3 in (C)]. Input was loaded to lane 1 as loading control. The experiment was repeated at least twice, with representative results shown here. ( D ) Heatmaps showing the SS18::SSX CUT&Tag signal densities in either HSSY II, SYO-1, or U2OS cells, ±5 kb from the centers of SS18::SSX peaks in HSSY II cells. ( E and F ) Venn diagram using the SS18::SSX peaks (E) or WDR5 peaks (F) called in HSSY II and SYO-1 cells. ( G ) Heatmaps showing the WDR5, SS18::SSX, and SMARCC1/BAF155 CUT&Tag signal intensities, ±5 kb from the centers of WDR5 peaks, in HSSY II cells. ( H and I ) Venn diagram showing overlap of SS18::SSX with the WDR5 and/or SMARCC1/BAF155 peaks in HSSY II (H) or SYO-1 (I) cells. ( J and K ) Genomic annotation (J) and gene ontology (GO) analysis (K) using the peaks cobound by SS18::SSX, WDR5, and SMARCC1/BAF155 in HSSY II cells as defined in (H). Y axis in (K) shows the −log 10 value of binomial P values.

Journal: Science Advances

Article Title: Pharmacologic degradation of WDR5 suppresses oncogenic activities of SS18::SSX and provides a therapeutic of synovial sarcoma

doi: 10.1126/sciadv.ads7876

Figure Lengend Snippet: ( A ) Representative co-IF images of SS18::SSX, WDR5 and DNA [probed with 4′,6-diamidino-2-phenylindole (DAPI)] in HSSY II cells, treated with 2.5 μM of DMSO (top) or MS67 (bottom) for 48 hours. Scale bar, 5 μm. The experiment was repeated at least twice, with representative results shown here. ( B and C ) co-IP to detect interaction between WDR and the SWI/SNF complex components in HSSY II (B) or U2OS cells (C) using either nonspecific IgG (lane 2), anti-SS18::SSX [lane 3 in (B)], or anti-WDR5 antibody [lane 4 in (B) and lane 3 in (C)]. Input was loaded to lane 1 as loading control. The experiment was repeated at least twice, with representative results shown here. ( D ) Heatmaps showing the SS18::SSX CUT&Tag signal densities in either HSSY II, SYO-1, or U2OS cells, ±5 kb from the centers of SS18::SSX peaks in HSSY II cells. ( E and F ) Venn diagram using the SS18::SSX peaks (E) or WDR5 peaks (F) called in HSSY II and SYO-1 cells. ( G ) Heatmaps showing the WDR5, SS18::SSX, and SMARCC1/BAF155 CUT&Tag signal intensities, ±5 kb from the centers of WDR5 peaks, in HSSY II cells. ( H and I ) Venn diagram showing overlap of SS18::SSX with the WDR5 and/or SMARCC1/BAF155 peaks in HSSY II (H) or SYO-1 (I) cells. ( J and K ) Genomic annotation (J) and gene ontology (GO) analysis (K) using the peaks cobound by SS18::SSX, WDR5, and SMARCC1/BAF155 in HSSY II cells as defined in (H). Y axis in (K) shows the −log 10 value of binomial P values.

Techniques: Co-Immunoprecipitation Assay, Control

$( A and B ) Heatmaps (A) and box plot of averaged intensities (B) of the indicated CUT&Tag or CUT&RUN signals (after normalization to spike-in control) of WDR5, SS18::SSX, or SMARCC1/BAF155 (±5 kb from the peak centers) in HSSY II cells, treated with 2.5 μM of DMSO or MS67 for 4 days. Wilcoxon test was used to generate P value. ( C and D ) Integrative Genomics Viewer (IGV) views of the indicated CUT&Tag or CUT&RUN signals at MNX1 (C) and SOX8 (D) in HSSY II cells, treated as in (A), or transduced with a control (shCtrl) or SS18::SSX-targeting shRNA (shSSX). ( E ) WB of SWI/SNF complex components using SYO-1 cells, treated with the indicated concentration of drugs for 48 hours. ( F and G ) WB of the indicated protein in the soluble nucleoplasmic (left) or chromatin-bound (right) fraction of HSSY II (F) or U2OS (G) cells, treated with 2.5 μM of DMSO or MS67 for 48 hours. Quantification of protein levels in (F) was normalized to soluble-DMSO fraction samples and labeled under the gel image. ( H ) Co-IP for interaction between WDR5 and the SS18::SSX-containing SWI/SNF complexes in HSSY II cells, first treated with 2.5 μM of DMSO (lanes 1 to 4) or MS67 (lanes 5 to 8) for 48 hours and then subject to IP using either nonspecific IgG, anti-WDR5, or anti-SS18::SSX antibody. WB and co-IP were repeated at least twice, with representative results shown here. ( I and J ) Heatmaps (I) and averaged intensities (J) of CUT&Tag signals (normalized to spike-in controls) of SS18::SSX and WDR5 (±5 kb from the peak centers) in HSSY II cells, transduced with the control (shCtrl) or SSX-targeting shRNA (shSSX). The y axis in (J) represents the average CUT&Tag signals and Wilcox test was used to generate P value.$

Journal: Science Advances

Article Title: Pharmacologic degradation of WDR5 suppresses oncogenic activities of SS18::SSX and provides a therapeutic of synovial sarcoma

doi: 10.1126/sciadv.ads7876

Figure Lengend Snippet: ( A and B ) Heatmaps (A) and box plot of averaged intensities (B) of the indicated CUT&Tag or CUT&RUN signals (after normalization to spike-in control) of WDR5, SS18::SSX, or SMARCC1/BAF155 (±5 kb from the peak centers) in HSSY II cells, treated with 2.5 μM of DMSO or MS67 for 4 days. Wilcoxon test was used to generate P value. ( C and D ) Integrative Genomics Viewer (IGV) views of the indicated CUT&Tag or CUT&RUN signals at MNX1 (C) and SOX8 (D) in HSSY II cells, treated as in (A), or transduced with a control (shCtrl) or SS18::SSX-targeting shRNA (shSSX). ( E ) WB of SWI/SNF complex components using SYO-1 cells, treated with the indicated concentration of drugs for 48 hours. ( F and G ) WB of the indicated protein in the soluble nucleoplasmic (left) or chromatin-bound (right) fraction of HSSY II (F) or U2OS (G) cells, treated with 2.5 μM of DMSO or MS67 for 48 hours. Quantification of protein levels in (F) was normalized to soluble-DMSO fraction samples and labeled under the gel image. ( H ) Co-IP for interaction between WDR5 and the SS18::SSX-containing SWI/SNF complexes in HSSY II cells, first treated with 2.5 μM of DMSO (lanes 1 to 4) or MS67 (lanes 5 to 8) for 48 hours and then subject to IP using either nonspecific IgG, anti-WDR5, or anti-SS18::SSX antibody. WB and co-IP were repeated at least twice, with representative results shown here. ( I and J ) Heatmaps (I) and averaged intensities (J) of CUT&Tag signals (normalized to spike-in controls) of SS18::SSX and WDR5 (±5 kb from the peak centers) in HSSY II cells, transduced with the control (shCtrl) or SSX-targeting shRNA (shSSX). The y axis in (J) represents the average CUT&Tag signals and Wilcox test was used to generate P value.

Techniques: Control, Transduction, shRNA, Concentration Assay, Labeling, Co-Immunoprecipitation Assay

( A ) Immunoblotting of H3K4me1, H3K4me2, and H3K4me3 in HSSY II (top) and SYO-1 cells (bottom), treated with the indicated concentration of DMSO, MS67, MS67N1, or MS67N2 for 48 hours. WB experiments above were repeated at least twice, with representative results shown here. ( B to E ) Heatmaps [(B) and (D)] and averaged intensities [(C) and (E)] of WDR5, H3K4me2, and H3K4me3 CUT&Tag signals after normalization to the spike-in control signals, ±5 kb from the centers of the called peaks, in HSSY II [(B) and (C)] and SYO-1 [(D) and (E)] cells treated by 2.5 μM of DMSO or MS67 for 4 days. ( F and G ) IGV views of the indicated developmental and stemness genes (F), as well as RP genes (G), in the DMSO (MS67 − )– or MS67 (MS67 + )–treated HSSY II (left) and SYO-1 cells (right).

Journal: Science Advances

Article Title: Pharmacologic degradation of WDR5 suppresses oncogenic activities of SS18::SSX and provides a therapeutic of synovial sarcoma

doi: 10.1126/sciadv.ads7876

Figure Lengend Snippet: ( A ) Immunoblotting of H3K4me1, H3K4me2, and H3K4me3 in HSSY II (top) and SYO-1 cells (bottom), treated with the indicated concentration of DMSO, MS67, MS67N1, or MS67N2 for 48 hours. WB experiments above were repeated at least twice, with representative results shown here. ( B to E ) Heatmaps [(B) and (D)] and averaged intensities [(C) and (E)] of WDR5, H3K4me2, and H3K4me3 CUT&Tag signals after normalization to the spike-in control signals, ±5 kb from the centers of the called peaks, in HSSY II [(B) and (C)] and SYO-1 [(D) and (E)] cells treated by 2.5 μM of DMSO or MS67 for 4 days. ( F and G ) IGV views of the indicated developmental and stemness genes (F), as well as RP genes (G), in the DMSO (MS67 − )– or MS67 (MS67 + )–treated HSSY II (left) and SYO-1 cells (right).

Techniques: Western Blot, Concentration Assay, Control

( A ) Venn diagram of DEGs in HSSY II or SYO-1 cells, down-regulated after treatment with 2.5 μM of MS67 versus DMSO for 2 or 4 days (d2 or d4). DEG is defined by a cutoff of fold-change (FC) more than 1.50 and P -adj less than 0.05. ( B to E ) Summary plot of GSEA [(B) and (C)] and the example enrichment for indicated pathways [(D) and (E)] in HSSY II or SYO-1 cells. The SS18::SSX-associated signature genes were defined to be those down-regulated significantly after SS18::SSX KD in HSSY II cells, with a cutoff of log 2 value of FC less than −1 and P -adj less than 0.001 using a dataset published in . ( F and G ) Box plots showing overall expression of the SS18::SSX signature genes, with Wilcoxon test used for generating P value. ( H ) Heatmap showing down-regulation of the indicated SS oncogenes in HSSY II (top) and SYO-1 cells (bottom), using the d2 treatment dataset. ( I and J ) RT-qPCR for the indicated gene in HSSY II (I) or SYO-1 cells (J), treated with 2.5 μM of DMSO or MS67 for 2 days ( n = 3; means ± SEM). P values were calculated with two-tail Student’s t test. ( K ) Venn diagram using MS67–down-regulated DEGs using the d2 and d4 treatment data and genes cobound by WDR5, SS18::SSX, and BAF155 in HSSY II cells. ( L ) Venn diagram using DEGs in HSSY II cells down-regulated because of stable transduction of shWDR5 versus control (HSSY II_shWDR5) or treatment of MS67 versus DMSO for 2 or 4 days (HSSY II_MS67_d2 or HSSY II_MS67_d4). ( M ) Heatmap showing down-regulation of the indicated SS oncogenes in HSSY II cells, stably transduced with the control (shCtrl-1 and shCtrl-2) or WDR5-targeting shRNA (shWDR5-1 or shWDR5-2). NES, normalized enrichment score; FDR, false discovery rate.

Journal: Science Advances

Article Title: Pharmacologic degradation of WDR5 suppresses oncogenic activities of SS18::SSX and provides a therapeutic of synovial sarcoma

doi: 10.1126/sciadv.ads7876

Figure Lengend Snippet: ( A ) Venn diagram of DEGs in HSSY II or SYO-1 cells, down-regulated after treatment with 2.5 μM of MS67 versus DMSO for 2 or 4 days (d2 or d4). DEG is defined by a cutoff of fold-change (FC) more than 1.50 and P -adj less than 0.05. ( B to E ) Summary plot of GSEA [(B) and (C)] and the example enrichment for indicated pathways [(D) and (E)] in HSSY II or SYO-1 cells. The SS18::SSX-associated signature genes were defined to be those down-regulated significantly after SS18::SSX KD in HSSY II cells, with a cutoff of log 2 value of FC less than −1 and P -adj less than 0.001 using a dataset published in . ( F and G ) Box plots showing overall expression of the SS18::SSX signature genes, with Wilcoxon test used for generating P value. ( H ) Heatmap showing down-regulation of the indicated SS oncogenes in HSSY II (top) and SYO-1 cells (bottom), using the d2 treatment dataset. ( I and J ) RT-qPCR for the indicated gene in HSSY II (I) or SYO-1 cells (J), treated with 2.5 μM of DMSO or MS67 for 2 days ( n = 3; means ± SEM). P values were calculated with two-tail Student’s t test. ( K ) Venn diagram using MS67–down-regulated DEGs using the d2 and d4 treatment data and genes cobound by WDR5, SS18::SSX, and BAF155 in HSSY II cells. ( L ) Venn diagram using DEGs in HSSY II cells down-regulated because of stable transduction of shWDR5 versus control (HSSY II_shWDR5) or treatment of MS67 versus DMSO for 2 or 4 days (HSSY II_MS67_d2 or HSSY II_MS67_d4). ( M ) Heatmap showing down-regulation of the indicated SS oncogenes in HSSY II cells, stably transduced with the control (shCtrl-1 and shCtrl-2) or WDR5-targeting shRNA (shWDR5-1 or shWDR5-2). NES, normalized enrichment score; FDR, false discovery rate.

Techniques: Expressing, Quantitative RT-PCR, Transduction, Control, Stable Transfection, shRNA

( A to C ) Growth of HSSY II CDX in NSG mice via subcutaneous (sc) inoculation [(A) means ± SEM in y axis], Kaplan-Meier survival curve (B), and averaged body weight of mice (C), treated with vehicle ( n = 7) or MS67 ( n = 7; a dose of 75 mg/kg, twice daily via intraperitoneal injection for 5 days per week). Statistical analysis for tumor growth was performed using two-way analysis of variance (ANOVA) followed by Sidak’s multiple comparison test, with statistical calculation at the last dosing time point labeled, while statistical analysis of survival was conducted by log-rank (Mantel-Cox) test. ( D to F ) Mass spectrometry–based measurement of MS67 concentration in the plasma and tumor samples (D), and WB (E) and RT-qPCR (F) for the indicated factor in tumors, collected 2 hours after the last dosing from vehicle- or MS67-treated mice. The numbers under gel images in (E) show the relative protein levels normalized to the Vehicle#1 sample. Y axis in (F) shows fold change of expression in the tumors from the MS67-treated versus vehicle-treated mice ( n = 3, means ± SEM). The two-tailed Student’s t test was used to calculate P value. ( G and H ) Representative IF images [scale bar, (G) 50 μm] and quantification of the percentage of cells positive for the indicated marker (H) in tissue sections of tumors, isolated from vehicle- or MS67-treated mice ( n = 5, means ± SEM). The two-tailed Student’s t test was used to calculate P value. ( I ) A model that the WDR5-containing KMT2/MLL complexes and the SS18::SSX-harboring SWI/SNF chromatin-remodeling complexes interact within the condensates and colocalize genome-wide, which operate to maintain both the openness and high H3K4 methylation levels at their target chromatin (demarcated by H2AK119ub, a histone modification recognized by a SSX segment in SS18::SSX), enforcing an oncogenic gene-expression program and SS pathogenesis.

Journal: Science Advances

Article Title: Pharmacologic degradation of WDR5 suppresses oncogenic activities of SS18::SSX and provides a therapeutic of synovial sarcoma

doi: 10.1126/sciadv.ads7876

Figure Lengend Snippet: ( A to C ) Growth of HSSY II CDX in NSG mice via subcutaneous (sc) inoculation [(A) means ± SEM in y axis], Kaplan-Meier survival curve (B), and averaged body weight of mice (C), treated with vehicle ( n = 7) or MS67 ( n = 7; a dose of 75 mg/kg, twice daily via intraperitoneal injection for 5 days per week). Statistical analysis for tumor growth was performed using two-way analysis of variance (ANOVA) followed by Sidak’s multiple comparison test, with statistical calculation at the last dosing time point labeled, while statistical analysis of survival was conducted by log-rank (Mantel-Cox) test. ( D to F ) Mass spectrometry–based measurement of MS67 concentration in the plasma and tumor samples (D), and WB (E) and RT-qPCR (F) for the indicated factor in tumors, collected 2 hours after the last dosing from vehicle- or MS67-treated mice. The numbers under gel images in (E) show the relative protein levels normalized to the Vehicle#1 sample. Y axis in (F) shows fold change of expression in the tumors from the MS67-treated versus vehicle-treated mice ( n = 3, means ± SEM). The two-tailed Student’s t test was used to calculate P value. ( G and H ) Representative IF images [scale bar, (G) 50 μm] and quantification of the percentage of cells positive for the indicated marker (H) in tissue sections of tumors, isolated from vehicle- or MS67-treated mice ( n = 5, means ± SEM). The two-tailed Student’s t test was used to calculate P value. ( I ) A model that the WDR5-containing KMT2/MLL complexes and the SS18::SSX-harboring SWI/SNF chromatin-remodeling complexes interact within the condensates and colocalize genome-wide, which operate to maintain both the openness and high H3K4 methylation levels at their target chromatin (demarcated by H2AK119ub, a histone modification recognized by a SSX segment in SS18::SSX), enforcing an oncogenic gene-expression program and SS pathogenesis.

Techniques: Injection, Comparison, Labeling, Mass Spectrometry, Concentration Assay, Clinical Proteomics, Quantitative RT-PCR, Expressing, Two Tailed Test, Marker, Isolation, Genome Wide, Methylation, Modification, Gene Expression

FIGURE 2 | Identification of methyltransferase mixed-lineage leukemia 1 (MLL1)/WD-40 repeat protein 5 (WDR5) and cellular senescence marker p16INK4a in human peritoneal mesothelial cells (HPMCs) from patients undergoing peritoneal dialysis (PD). (A) Representative western blotting analyses showing MLL1, WDR5, H3K4 trimethylation (H3K4me3), and p16INK4a expression in HPMCs from non-PD and PD patients. (B) Quantitative analysis of MLL1, WDR5, H3K4me3, and p16INK4a protein expression in HPMCs from non-PD and PD. Protein abundance for MLL1, WDR5, and p16INK4a was normalized to that of α-tubulin, and H3K4me3 protein abundance was normalized to that of histone H3 (H3). n = 5 per group. *p < 0.05, and **p < 0.01 by two-tailed Student's t test. (C) Correlation analysis between peritoneal function (D/P Cre) and p16INK4a expression in HPMCs from patients undergoing PD (n = 10) using Spearman's rank correlation test. ρ, Spearman correlation coefficient; ρ = 0.82, p = 0.003.

Journal: The FASEB Journal

Article Title: Targeting MLL1/WDR5‐Mediated Epigenetic Regulation Mitigates Peritoneal Fibrosis by Reducing p16^INK4a

doi: 10.1096/fj.202402382r

Figure Lengend Snippet: FIGURE 2 | Identification of methyltransferase mixed-lineage leukemia 1 (MLL1)/WD-40 repeat protein 5 (WDR5) and cellular senescence marker p16INK4a in human peritoneal mesothelial cells (HPMCs) from patients undergoing peritoneal dialysis (PD). (A) Representative western blotting analyses showing MLL1, WDR5, H3K4 trimethylation (H3K4me3), and p16INK4a expression in HPMCs from non-PD and PD patients. (B) Quantitative analysis of MLL1, WDR5, H3K4me3, and p16INK4a protein expression in HPMCs from non-PD and PD. Protein abundance for MLL1, WDR5, and p16INK4a was normalized to that of α-tubulin, and H3K4me3 protein abundance was normalized to that of histone H3 (H3). n = 5 per group. *p < 0.05, and **p < 0.01 by two-tailed Student's t test. (C) Correlation analysis between peritoneal function (D/P Cre) and p16INK4a expression in HPMCs from patients undergoing PD (n = 10) using Spearman's rank correlation test. ρ, Spearman correlation coefficient; ρ = 0.82, p = 0.003.

Article Snippet: The following antibodies were used: rabbit monoclonal anti- MLL1 antibody (1:1000, #14197, Cell Signaling Technology, Danvers, MA, USA, RRID:AB_2688010), rabbit monoclonal anti- WDR5 (1:1000, #13105, Cell Signaling Technology, RRID:AB_2620133), rabbit polyclonal antiH3K4me3 (1:2000, ab8580, Abcam, RRID:AB_306649), rabbit monoclonal anti- p16INK4a (1:2500, ab51243, Abcam, RRID:AB_2059963), rabbit monoclonal anti- p21WAF1/CIP1 (1:1000, ab109199, Abcam, RRID: AB_10861551), rabbit polyclonal anti- p53 (1:1000, #9282, Cell Signaling Technology, RRID: AB_331476), mouse monoclonal anti- α- SMA (1:5000, A2547, Sigma- Aldrich, St. Louis, MO, USA, RRID:AB_476701), rabbit polyclonal anti- collagen Iα1 (1:1000, NBP1- 30054, 15306860, 2025, 8, D ow nloaded from https://faseb.onlinelibrary.w iley.com /doi/10.1096/fj.202402382R , W iley O nline L ibrary on [22/05/2025].

Techniques: Marker, Western Blot, Expressing, Quantitative Proteomics, Two Tailed Test

FIGURE 3 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 repeat protein 5 (WDR5) complex inhibitors on H3K4 trimethylation (H3K4me3) and p16INK4a expression in mice with peritoneal fibrosis. (A) Representative hematoxylin–eosin staining of peritoneal tissues from control mice and methylglyoxal (MGO)-injected mice. Bars indicate the thickness of submesothelial compact zone (scale bar: 100 μm). (B) Representative immunohis- tochemical analyses showing the expression of MLL1 and WDR5 in peritoneal tissues from control mice and MGO-injected mice. The right panels show high-magnification images of the boxed regions (scale bar: 100 μm). (C) Quantitative analysis of MLL1- and WDR5-positive cells in peritoneal tissues. ***p < 0.001 by 2-tailed Student's t test. (D, F) Representative immunohistochemical analyses showing H3K4me3 and p16INK4a expression in peritoneal tissue from control mice, MGO-injected mice, and MGO-injected mice receiving (D) MM-102 and (F) OICR-9429. The right panels show high-magnification images of the boxed regions (scale bar: 100 μm). (E, G) Quantitative analysis of H3K4me3-positive cells and p16INK4a-positive cells in peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice receiving (E) MM-102 and (G) OICR-9429. n = 5 per group. **p < 0.01 and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Journal: The FASEB Journal

Article Title: Targeting MLL1/WDR5‐Mediated Epigenetic Regulation Mitigates Peritoneal Fibrosis by Reducing p16^INK4a

doi: 10.1096/fj.202402382r

Figure Lengend Snippet: FIGURE 3 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 repeat protein 5 (WDR5) complex inhibitors on H3K4 trimethylation (H3K4me3) and p16INK4a expression in mice with peritoneal fibrosis. (A) Representative hematoxylin–eosin staining of peritoneal tissues from control mice and methylglyoxal (MGO)-injected mice. Bars indicate the thickness of submesothelial compact zone (scale bar: 100 μm). (B) Representative immunohis- tochemical analyses showing the expression of MLL1 and WDR5 in peritoneal tissues from control mice and MGO-injected mice. The right panels show high-magnification images of the boxed regions (scale bar: 100 μm). (C) Quantitative analysis of MLL1- and WDR5-positive cells in peritoneal tissues. ***p < 0.001 by 2-tailed Student's t test. (D, F) Representative immunohistochemical analyses showing H3K4me3 and p16INK4a expression in peritoneal tissue from control mice, MGO-injected mice, and MGO-injected mice receiving (D) MM-102 and (F) OICR-9429. The right panels show high-magnification images of the boxed regions (scale bar: 100 μm). (E, G) Quantitative analysis of H3K4me3-positive cells and p16INK4a-positive cells in peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice receiving (E) MM-102 and (G) OICR-9429. n = 5 per group. **p < 0.01 and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Techniques: Expressing, Staining, Control, Injection, Immunohistochemical staining

FIGURE 4 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 repeat protein 5 (WDR5) complex inhibitors on chronic peritoneal injury following methylglyoxal (MGO) administration. (A, C) Representative hematoxylin–eosin staining and Masson's trichrome staining of peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice treated with (A) MM-102 and (C) OICR-9429. The right panels show high- magnification images of the boxed regions (scale bar: 200 μm). Bars indicate the thickness of the submesothelial compact zone. (B, D) Quantitative analysis of cell density and submesothelial compact zone thickness in peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice treated with (B) MM-102 and (D) OICR-9429. n = 5 per group. **p < 0.01 and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Journal: The FASEB Journal

Article Title: Targeting MLL1/WDR5‐Mediated Epigenetic Regulation Mitigates Peritoneal Fibrosis by Reducing p16^INK4a

doi: 10.1096/fj.202402382r

Figure Lengend Snippet: FIGURE 4 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 repeat protein 5 (WDR5) complex inhibitors on chronic peritoneal injury following methylglyoxal (MGO) administration. (A, C) Representative hematoxylin–eosin staining and Masson's trichrome staining of peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice treated with (A) MM-102 and (C) OICR-9429. The right panels show high- magnification images of the boxed regions (scale bar: 200 μm). Bars indicate the thickness of the submesothelial compact zone. (B, D) Quantitative analysis of cell density and submesothelial compact zone thickness in peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice treated with (B) MM-102 and (D) OICR-9429. n = 5 per group. **p < 0.01 and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Techniques: Staining, Control, Injection

FIGURE 5 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 repeat protein 5 (WDR5) complex inhibitors on methylglyoxal (MGO)-induced peritoneal fibrosis in mice. (A, D) mRNA abundance of Acta2 and Col1a1 in control mice, MGO-injected mice, and MGO-injected mice treated with (A) MM-102 and (D) OICR-9429. Gene expression was normalized to the internal control Gapdh. n = 5 per group. (B, E) Representative immunohis- tochemical analyses showing α-smooth muscle actin (SMA), fibroblast-specific protein (FSP)-1, and collagen I expression in peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice treated with (B) MM-102 and (E) OICR-9429. The right panels show high-magnification images of the boxed regions (scale bar: 100 μm). (C, F) Quantitative analysis of α-SMA-positive cells, FSP-1-positive cells, and collagen I-positive areas in peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice treated with (C) MM-102 and (E) OICR-9429. n = 5 per group. **p < 0.01, and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Journal: The FASEB Journal

Article Title: Targeting MLL1/WDR5‐Mediated Epigenetic Regulation Mitigates Peritoneal Fibrosis by Reducing p16^INK4a

doi: 10.1096/fj.202402382r

Figure Lengend Snippet: FIGURE 5 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 repeat protein 5 (WDR5) complex inhibitors on methylglyoxal (MGO)-induced peritoneal fibrosis in mice. (A, D) mRNA abundance of Acta2 and Col1a1 in control mice, MGO-injected mice, and MGO-injected mice treated with (A) MM-102 and (D) OICR-9429. Gene expression was normalized to the internal control Gapdh. n = 5 per group. (B, E) Representative immunohis- tochemical analyses showing α-smooth muscle actin (SMA), fibroblast-specific protein (FSP)-1, and collagen I expression in peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice treated with (B) MM-102 and (E) OICR-9429. The right panels show high-magnification images of the boxed regions (scale bar: 100 μm). (C, F) Quantitative analysis of α-SMA-positive cells, FSP-1-positive cells, and collagen I-positive areas in peritoneal tissues from control mice, MGO-injected mice, and MGO-injected mice treated with (C) MM-102 and (E) OICR-9429. n = 5 per group. **p < 0.01, and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Techniques: Control, Injection, Gene Expression, Expressing

FIGURE 6 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 repeat protein 5 (WDR5) complex inhibitors on peritoneal inflammation. (A, D) mRNA abundance of Il1b and Tnf in control mice, methylglyoxal (MGO)-injected mice, and MGO-injected mice treated with (A) MM-102 and (D) OICR-9429. Gene expression was normalized to the internal control Gapdh. n = 5 per group. (B, E) Representative immunohistochemical analyses showing interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α expression in peritoneal tissue from control mice, MGO-injected mice, and MGO-injected mice treated with (B) MM-102 and (E) OICR-9429. The right panels show high-magnification images of the boxed regions (scale bar: 100 μm). (C, F) Quantitative analysis of IL-1β-positive cells, IL-6-positive cells, and TNF-α-positive cells in peritoneal tissue from control mice, MGO- injected mice, and MGO-injected mice treated with (C) MM-102 and (F) OICR-9429. n = 5 per group. *p < 0.05, **p < 0.01, and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Journal: The FASEB Journal

Article Title: Targeting MLL1/WDR5‐Mediated Epigenetic Regulation Mitigates Peritoneal Fibrosis by Reducing p16^INK4a

doi: 10.1096/fj.202402382r

Figure Lengend Snippet: FIGURE 6 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 repeat protein 5 (WDR5) complex inhibitors on peritoneal inflammation. (A, D) mRNA abundance of Il1b and Tnf in control mice, methylglyoxal (MGO)-injected mice, and MGO-injected mice treated with (A) MM-102 and (D) OICR-9429. Gene expression was normalized to the internal control Gapdh. n = 5 per group. (B, E) Representative immunohistochemical analyses showing interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α expression in peritoneal tissue from control mice, MGO-injected mice, and MGO-injected mice treated with (B) MM-102 and (E) OICR-9429. The right panels show high-magnification images of the boxed regions (scale bar: 100 μm). (C, F) Quantitative analysis of IL-1β-positive cells, IL-6-positive cells, and TNF-α-positive cells in peritoneal tissue from control mice, MGO- injected mice, and MGO-injected mice treated with (C) MM-102 and (F) OICR-9429. n = 5 per group. *p < 0.05, **p < 0.01, and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Techniques: Control, Injection, Gene Expression, Immunohistochemical staining, Expressing

FIGURE 7 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 re- peat protein 5 (WDR5) complex inhibitors on functional impairment of the peritoneal membrane in mice with peritoneal fibrosis. (A, B) The dialysate/plasma ratio of urea nitrogen (D/P UN) and the peritoneal absorption of glucose from dialysate (D/D0 glucose) were assessed in control mice, methylglyoxal (MGO)-injected mice, and MGO-injected mice treated with (A) MM-102 and (B) OICR-9429 during a 10 min di- alysate dwell time (4.25% dialysis solution). n = 5 per group. **p < 0.01 and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Journal: The FASEB Journal

Article Title: Targeting MLL1/WDR5‐Mediated Epigenetic Regulation Mitigates Peritoneal Fibrosis by Reducing p16^INK4a

doi: 10.1096/fj.202402382r

Figure Lengend Snippet: FIGURE 7 | Effects of mixed-lineage leukemia 1 (MLL1)/ WD-40 re- peat protein 5 (WDR5) complex inhibitors on functional impairment of the peritoneal membrane in mice with peritoneal fibrosis. (A, B) The dialysate/plasma ratio of urea nitrogen (D/P UN) and the peritoneal absorption of glucose from dialysate (D/D0 glucose) were assessed in control mice, methylglyoxal (MGO)-injected mice, and MGO-injected mice treated with (A) MM-102 and (B) OICR-9429 during a 10 min di- alysate dwell time (4.25% dialysis solution). n = 5 per group. **p < 0.01 and ***p < 0.001 by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test. CTL, control.

Techniques: Functional Assay, Membrane, Clinical Proteomics, Control, Injection

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