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anti sumo1 (Danaher Inc)

Name: anti sumo1
Brand: Danaher Inc
Rating: 86 (1 reviews)

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Danaher Inc anti sumo1

Zinc deficiency reduced <t>SUMO1</t> expression in cartilage tissue. (A) Cartilage tissues from ZnD and ZnA groups were immunohistochemically stained with SUMO1 antibody (scale bar, 50 um, 40×). (B) Free and conjugated SUMO1 expression levels in ZnD and ZnA groups were analyzed by WB. The primary chondrocytes were used for WB assay. (C) Quantitative analysis of the WB results in B. * p < 0.05 compared with controls.

Anti Sumo1, supplied by Danaher 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 sumo1/product/Danaher Inc
Average 86 stars, based on 1 article reviews

anti sumo1 - by Bioz Stars, 2025-02

86/100 stars

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1) Product Images from "The Role of SUMO1 Modification of SOX9 in Cartilage Development Stimulated by Zinc Ions in Mice"

Article Title: The Role of SUMO1 Modification of SOX9 in Cartilage Development Stimulated by Zinc Ions in Mice

Journal: Organogenesis

doi: 10.1080/15476278.2025.2460269

Zinc deficiency reduced SUMO1 expression in cartilage tissue. (A) Cartilage tissues from ZnD and ZnA groups were immunohistochemically stained with SUMO1 antibody (scale bar, 50 um, 40×). (B) Free and conjugated SUMO1 expression levels in ZnD and ZnA groups were analyzed by WB. The primary chondrocytes were used for WB assay. (C) Quantitative analysis of the WB results in B. * p < 0.05 compared with controls.

Figure Legend Snippet: Zinc deficiency reduced SUMO1 expression in cartilage tissue. (A) Cartilage tissues from ZnD and ZnA groups were immunohistochemically stained with SUMO1 antibody (scale bar, 50 um, 40×). (B) Free and conjugated SUMO1 expression levels in ZnD and ZnA groups were analyzed by WB. The primary chondrocytes were used for WB assay. (C) Quantitative analysis of the WB results in B. * p < 0.05 compared with controls.

Techniques Used: Expressing, Staining

Zinc induced SUMOylation of SOX9. Immunoprecipitation-immunoblot analyses examined the conjugation between SOX9 and SUMO1. Compared with the control group, zinc stimulated SOX9 SUMOylation (A and B). (C) one Lys (K) residues were replaced with arginine (R) through point mutagenesis. Flag-tagged SOX9 WT and K – R Mut were transfected with HA-SUMO1 and His-UBC9 into ATDC5. The transfected cells were lysed in a denaturing buffer and diluted in a non-denaturing buffer. The lysates were immunoprecipitated with an anti-Flag antibody and examined by WB using the indicated antibodies (right).

Figure Legend Snippet: Zinc induced SUMOylation of SOX9. Immunoprecipitation-immunoblot analyses examined the conjugation between SOX9 and SUMO1. Compared with the control group, zinc stimulated SOX9 SUMOylation (A and B). (C) one Lys (K) residues were replaced with arginine (R) through point mutagenesis. Flag-tagged SOX9 WT and K – R Mut were transfected with HA-SUMO1 and His-UBC9 into ATDC5. The transfected cells were lysed in a denaturing buffer and diluted in a non-denaturing buffer. The lysates were immunoprecipitated with an anti-Flag antibody and examined by WB using the indicated antibodies (right).

Techniques Used: Immunoprecipitation, Western Blot, Conjugation Assay, Control, Mutagenesis, Transfection

Image Search Results

Journal: Organogenesis

Article Title: The Role of SUMO1 Modification of SOX9 in Cartilage Development Stimulated by Zinc Ions in Mice

doi: 10.1080/15476278.2025.2460269

Figure Lengend Snippet: Zinc deficiency reduced SUMO1 expression in cartilage tissue. (A) Cartilage tissues from ZnD and ZnA groups were immunohistochemically stained with SUMO1 antibody (scale bar, 50 um, 40×). (B) Free and conjugated SUMO1 expression levels in ZnD and ZnA groups were analyzed by WB. The primary chondrocytes were used for WB assay. (C) Quantitative analysis of the WB results in B. * p < 0.05 compared with controls.

Article Snippet: We incubated the membranes with primary antibodies (anti-SUMO1 (Abcam ab32058), anti-SUMO2/3 (Abcam ab81371), anti-UBC9 (CST #4918), anti-SOX9 (Abcam ab185230) anti-MMP13 (Abcam ab219620), anti-Collagen II (Abcam ab307674), anti-RUNX2 (Abcam ab236639), anti-aggrecan (Abcam ab3778)), and secondary antibodies (goat anti-mouse IgG-HRP (Abcam ab6789) or goat anti-rabbit IgG-HRP (Abcam ab6721)) sequentially before developing and exposing them.

Techniques: Expressing, Staining

Journal: Organogenesis

Article Title: The Role of SUMO1 Modification of SOX9 in Cartilage Development Stimulated by Zinc Ions in Mice

doi: 10.1080/15476278.2025.2460269

Figure Lengend Snippet: Zinc induced SUMOylation of SOX9. Immunoprecipitation-immunoblot analyses examined the conjugation between SOX9 and SUMO1. Compared with the control group, zinc stimulated SOX9 SUMOylation (A and B). (C) one Lys (K) residues were replaced with arginine (R) through point mutagenesis. Flag-tagged SOX9 WT and K – R Mut were transfected with HA-SUMO1 and His-UBC9 into ATDC5. The transfected cells were lysed in a denaturing buffer and diluted in a non-denaturing buffer. The lysates were immunoprecipitated with an anti-Flag antibody and examined by WB using the indicated antibodies (right).

Techniques: Immunoprecipitation, Western Blot, Conjugation Assay, Control, Mutagenesis, Transfection

Journal: Organogenesis

Article Title: The Role of SUMO1 Modification of SOX9 in Cartilage Development Stimulated by Zinc Ions in Mice

doi: 10.1080/15476278.2025.2460269

Article Snippet: We centrifuged the lysates and incubated the supernatants overnight with anti-SUMO1 antibody (Abcam ab32058) or anti-SOX9 antibody (Abcam ab185230).

Techniques: Expressing, Staining

Journal: Organogenesis

Article Title: The Role of SUMO1 Modification of SOX9 in Cartilage Development Stimulated by Zinc Ions in Mice

doi: 10.1080/15476278.2025.2460269

Article Snippet: We centrifuged the lysates and incubated the supernatants overnight with anti-SUMO1 antibody (Abcam ab32058) or anti-SOX9 antibody (Abcam ab185230).

Techniques: Immunoprecipitation, Western Blot, Conjugation Assay, Control, Mutagenesis, Transfection

(A) Representative images of PML KO cells expressing mCh-eDHFR-SUMO3, 3xHalo-TRF1, and PML(SUMO-) with the dimerizer to dimerize at indicated time points. (B) Quantification of APB and PML NB numbers per cell after adding the dimerizer when recruiting SUMO3 in PML KO cells expressing PML(SIM-) and (C) PML(SUMO-). (D) Representative images of PML KO cells expressing mCh-eDHFR-SIM, 3xHalo-TRF1, and PML(SIM-) with the dimerizer to dimerize at indicated time points. (E) Quantification of APB and PML NB numbers per cell after adding the dimerizer when recruiting SIM in PML KO cells expressing PML(SIM-) and (G) PML(SUMO-). Numbers in (B)(C)(E)(G) are normalized by the number at the first time point for each cell, more than 17 cells per group, three independent experiments. (F) Representative images of PML KO cells expressing mCh-eDHFR-SIM, 3xHalo-TRF1, and PML(SUMO-) with the dimerizer to dimerize at indicated time points. (H) Representative images and (I) quantification of SUMO1 localization on telomeres in G2 arrested PML KO after overexpressing PML WT, PML(SUMO-) or PML(SIM-). Each dot represents one cell in three independent experiments, more than 54 cells were analyzed in each group. (J) Number of APBs in G2 arrested PML KO after overexpressing PML WT, PML(SUMO-), or PML(SIM-). Each dot represents one cell in three independent experiments, more than 45 cells were analyzed in each group. Scale bars, 5 μm.

Journal: bioRxiv

Article Title: Telomeric SUMO level influences the choices of APB formation pathways and ALT efficiency

doi: 10.1101/2025.01.16.633463

Figure Lengend Snippet: (A) Representative images of PML KO cells expressing mCh-eDHFR-SUMO3, 3xHalo-TRF1, and PML(SUMO-) with the dimerizer to dimerize at indicated time points. (B) Quantification of APB and PML NB numbers per cell after adding the dimerizer when recruiting SUMO3 in PML KO cells expressing PML(SIM-) and (C) PML(SUMO-). (D) Representative images of PML KO cells expressing mCh-eDHFR-SIM, 3xHalo-TRF1, and PML(SIM-) with the dimerizer to dimerize at indicated time points. (E) Quantification of APB and PML NB numbers per cell after adding the dimerizer when recruiting SIM in PML KO cells expressing PML(SIM-) and (G) PML(SUMO-). Numbers in (B)(C)(E)(G) are normalized by the number at the first time point for each cell, more than 17 cells per group, three independent experiments. (F) Representative images of PML KO cells expressing mCh-eDHFR-SIM, 3xHalo-TRF1, and PML(SUMO-) with the dimerizer to dimerize at indicated time points. (H) Representative images and (I) quantification of SUMO1 localization on telomeres in G2 arrested PML KO after overexpressing PML WT, PML(SUMO-) or PML(SIM-). Each dot represents one cell in three independent experiments, more than 54 cells were analyzed in each group. (J) Number of APBs in G2 arrested PML KO after overexpressing PML WT, PML(SUMO-), or PML(SIM-). Each dot represents one cell in three independent experiments, more than 45 cells were analyzed in each group. Scale bars, 5 μm.

Article Snippet: The resulting whole-cell lysates were analyzed by Western blotting with the following primary antibodies: SUMO1 (Ab32058, Abcam,1:1000 dilution), SUMO2/3 (Asm23, Cytoskeleton, 1:1000 dilution), MMS21 (A304-129A, Bethyl, 1:1000 dilution), PIAS4 (4392, Cell Signaling, 1:1000 dilution), TRF2 (NBP1-86911, Novus, 1:1000 dilution), mCherry (ab183628, Abcam, 1:1000 dilution), BLM (A300-110A, Bethyl, 1:1000 dilution), HA (3724S, Cell signaling, 1:1000 dilution) and β-Actin (Ab8226, Abcam, 1:1000 dilution).

Techniques: Expressing

(A) Representative images and (B) quantification of SUMO1 localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 56 cells were analyzed in each group. (C) Quantification of SUMO2/3 localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 50 cells were analyzed in each group. (D) Quantification of telomere number in G2 arrested U2OS WT after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 56 cells were analyzed in each group. Scale bars, 5 μm.

Journal: bioRxiv

Article Title: Telomeric SUMO level influences the choices of APB formation pathways and ALT efficiency

doi: 10.1101/2025.01.16.633463

Figure Lengend Snippet: (A) Representative images and (B) quantification of SUMO1 localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 56 cells were analyzed in each group. (C) Quantification of SUMO2/3 localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 50 cells were analyzed in each group. (D) Quantification of telomere number in G2 arrested U2OS WT after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 56 cells were analyzed in each group. Scale bars, 5 μm.

Techniques: Transfection, Control

(A) Western blot of SUMO1, SUMO2/3 (same antibody), or β-Actin after transfection of control siRNA, siSUMO1 or siSUMO2/3 in WT U2OS and PML KO U2OS cells for two days. (B) Representative images of telomeres with EdU, PML and (E) SUMO2/3 staining in G2-arrested U2OS WT cells and PML KO cells after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Yellow arrows indicate EdU signals at telomeres. (C) Quantification for the number of PML NBs and APBs in G2 arrested U2OS WT cells after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 46 cells were analyzed in each group. (D) Quantification of the number of EdU foci on telomeres in G2 arrested U2OS WT and PML KO cells after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 50 cells were analyzed in each group. (F) Quantification of newly formed APBs per cell after FokI induction for 2 hours, with transfection of control siRNA, siSUMO1 or siSUMO2/3 for 2 days. More than 23 cells were analyzed in each group from three independent experiments. Scale bars, 5 μm.

Journal: bioRxiv

Article Title: Telomeric SUMO level influences the choices of APB formation pathways and ALT efficiency

doi: 10.1101/2025.01.16.633463

Figure Lengend Snippet: (A) Western blot of SUMO1, SUMO2/3 (same antibody), or β-Actin after transfection of control siRNA, siSUMO1 or siSUMO2/3 in WT U2OS and PML KO U2OS cells for two days. (B) Representative images of telomeres with EdU, PML and (E) SUMO2/3 staining in G2-arrested U2OS WT cells and PML KO cells after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Yellow arrows indicate EdU signals at telomeres. (C) Quantification for the number of PML NBs and APBs in G2 arrested U2OS WT cells after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 46 cells were analyzed in each group. (D) Quantification of the number of EdU foci on telomeres in G2 arrested U2OS WT and PML KO cells after transfection of control siRNA, siSUMO1 or siSUMO2/3 for two days. Each dot represents one cell in three independent experiments, more than 50 cells were analyzed in each group. (F) Quantification of newly formed APBs per cell after FokI induction for 2 hours, with transfection of control siRNA, siSUMO1 or siSUMO2/3 for 2 days. More than 23 cells were analyzed in each group from three independent experiments. Scale bars, 5 μm.

Techniques: Western Blot, Transfection, Control, Staining

(A) Representative images and (B) quantification of SUMO1 localization on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 52 cells were analyzed in each group. (C) Representative images and (D) quantification of SUMO2/3 localization on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 55 cells were analyzed in each group. (E) Representative images and (F) quantification of PML localization on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 50 cells were analyzed in each group. (G) Quantification of total PML NBs in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 50 cells were analyzed in each group. (H) Quantification of telomere number in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 53 cells were analyzed in each group. (I) Quantification of EdU on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 42 cells were analyzed in each group. (J) Representative images of EdU and PCNA or (L) RPA on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. (K) Quantification of endogenous PCNA and RPA localization on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one independent experiment, more than 22 cells were analyzed in each group for three independent experiments. (M) Western blot of SUMO1, SUMO2/3 or β-Actin in SUMO1 KO, SUMO2 KO, SUMO2 KO+SUMO2, and control U2OS cells. Scale bars, 5 μm.

Journal: bioRxiv

Article Title: Telomeric SUMO level influences the choices of APB formation pathways and ALT efficiency

doi: 10.1101/2025.01.16.633463

Figure Lengend Snippet: (A) Representative images and (B) quantification of SUMO1 localization on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 52 cells were analyzed in each group. (C) Representative images and (D) quantification of SUMO2/3 localization on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 55 cells were analyzed in each group. (E) Representative images and (F) quantification of PML localization on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 50 cells were analyzed in each group. (G) Quantification of total PML NBs in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 50 cells were analyzed in each group. (H) Quantification of telomere number in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 53 cells were analyzed in each group. (I) Quantification of EdU on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one cell in three independent experiments, more than 42 cells were analyzed in each group. (J) Representative images of EdU and PCNA or (L) RPA on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. (K) Quantification of endogenous PCNA and RPA localization on telomeres in G2 arrested U2OS WT, SUMO1 KO, SUMO2 KO, and SUMO2 KO+SUMO2 cells. Each dot represents one independent experiment, more than 22 cells were analyzed in each group for three independent experiments. (M) Western blot of SUMO1, SUMO2/3 or β-Actin in SUMO1 KO, SUMO2 KO, SUMO2 KO+SUMO2, and control U2OS cells. Scale bars, 5 μm.

Techniques: Western Blot, Control

(A) Representative images and (B) quantification of PML localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one cell in three independent experiments, more than 45 cells were analyzed in each group. (C) Quantification of total PML NBs in G2 arrested U2OS WT after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one cell in three independent experiments, more than 46 cells were analyzed in each group. (D) Quantification of SUMO1 localization on telomeres and (E) telomere number in G2 arrested U2OS WT and PML KO cells after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one cell in three independent experiments, more than 53 cells were analyzed in each group. (F) Representative images and (G) quantification of PCNA localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one independent experiment, more than 23 cells were analyzed in each group for three independent experiments. (H) Representative images and (I) quantification of RPA localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one independent experiment, more than 28 cells were analyzed in each group for three independent experiments. Scale bars, 5 μm.

Journal: bioRxiv

Article Title: Telomeric SUMO level influences the choices of APB formation pathways and ALT efficiency

doi: 10.1101/2025.01.16.633463

Figure Lengend Snippet: (A) Representative images and (B) quantification of PML localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one cell in three independent experiments, more than 45 cells were analyzed in each group. (C) Quantification of total PML NBs in G2 arrested U2OS WT after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one cell in three independent experiments, more than 46 cells were analyzed in each group. (D) Quantification of SUMO1 localization on telomeres and (E) telomere number in G2 arrested U2OS WT and PML KO cells after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one cell in three independent experiments, more than 53 cells were analyzed in each group. (F) Representative images and (G) quantification of PCNA localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one independent experiment, more than 23 cells were analyzed in each group for three independent experiments. (H) Representative images and (I) quantification of RPA localization on telomeres in G2 arrested U2OS WT after transfection of control siRNA, siMMS21, or siPIAS4 for two days. Each dot represents one independent experiment, more than 28 cells were analyzed in each group for three independent experiments. Scale bars, 5 μm.

Techniques: Transfection, Control

(A) Representative images and (B) quantification of SUMO1 localization at telomeres in U2OS WT and PML KO cells after dimerizing MMS21(C215A)/PIAS4(C215A) to telomeres for 6 hours. Each dot represents one cell in three independent experiments, more than 54 cells were analyzed in each group. (C) Representative images and (D) quantification of SUMO2/3 localization at telomeres in U2OS WT and PML KO cells after dimerizing MMS21 or PIAS4 to telomeres for 6 hours. Each dot represents one cell in three independent experiments, more than 54 cells were analyzed in each group. (E) Representative images of PML localization on telomeres in U2OS WT after dimerizing PIAS4, PIAS4(C342A), Rad52-PIAS4, or Rad52-PIAS4(C342A) to telomeres for 6 hours. (F) Quantification of telomere number in U2OS WT and PML KO cells after dimerizing MMS21, PIAS4, MMS21(C215A), or PIAS4(C215A) to telomeres for 6 hours. Each dot represents one cell in three independent experiments, more than 55 cells were analyzed in each group. (G) Quantification of SUMO1/2/3 intensity per unit of MMS21 or PIAS4 intensity on telomeres in U2OS WT and PML KO cells. Each dot represents one cell in three independent experiments, more than 52 cells were analyzed in each group. (H) Quantification of telomere number in U2OS WT cells after dimerizing MMS21/PIAS4/Rad52-MMS21(C215A)/Rad52-PIAS4(C342A) to telomeres for 6 hours. Each dot represents one cell in three independent experiments, more than 46 cells were analyzed in each group. (I) U2OS cells expressing Halo-GFP-TRF1, HA-SUMO3, mCherry-eDHFR-MMS21(C215A), or (J) mCherry-eDHFR-PIAS4(C342A) with adding 5-hour of dimerizer. SUMOylated proteins were captured with protein A/G beads following HA antibody incubation. Levels of TRF2 and HA in inputs and HA pull-downs were analyzed by western blot. Scale bars, 5 μm.

Journal: bioRxiv

Article Title: Telomeric SUMO level influences the choices of APB formation pathways and ALT efficiency

doi: 10.1101/2025.01.16.633463

Figure Lengend Snippet: (A) Representative images and (B) quantification of SUMO1 localization at telomeres in U2OS WT and PML KO cells after dimerizing MMS21(C215A)/PIAS4(C215A) to telomeres for 6 hours. Each dot represents one cell in three independent experiments, more than 54 cells were analyzed in each group. (C) Representative images and (D) quantification of SUMO2/3 localization at telomeres in U2OS WT and PML KO cells after dimerizing MMS21 or PIAS4 to telomeres for 6 hours. Each dot represents one cell in three independent experiments, more than 54 cells were analyzed in each group. (E) Representative images of PML localization on telomeres in U2OS WT after dimerizing PIAS4, PIAS4(C342A), Rad52-PIAS4, or Rad52-PIAS4(C342A) to telomeres for 6 hours. (F) Quantification of telomere number in U2OS WT and PML KO cells after dimerizing MMS21, PIAS4, MMS21(C215A), or PIAS4(C215A) to telomeres for 6 hours. Each dot represents one cell in three independent experiments, more than 55 cells were analyzed in each group. (G) Quantification of SUMO1/2/3 intensity per unit of MMS21 or PIAS4 intensity on telomeres in U2OS WT and PML KO cells. Each dot represents one cell in three independent experiments, more than 52 cells were analyzed in each group. (H) Quantification of telomere number in U2OS WT cells after dimerizing MMS21/PIAS4/Rad52-MMS21(C215A)/Rad52-PIAS4(C342A) to telomeres for 6 hours. Each dot represents one cell in three independent experiments, more than 46 cells were analyzed in each group. (I) U2OS cells expressing Halo-GFP-TRF1, HA-SUMO3, mCherry-eDHFR-MMS21(C215A), or (J) mCherry-eDHFR-PIAS4(C342A) with adding 5-hour of dimerizer. SUMOylated proteins were captured with protein A/G beads following HA antibody incubation. Levels of TRF2 and HA in inputs and HA pull-downs were analyzed by western blot. Scale bars, 5 μm.

Techniques: Expressing, Incubation, Western Blot

(A) Representative images and (B) quantification of SUMO1/2/3 intensity per unit of MMS21/PIAS4 or Rad52-MMS21/PIAS4/MMS21(C215A)/PIAS4(C342A) intensity on telomeres in U2OS cells. Each dot represents one cell in three independent experiments, more than 47 cells were analyzed in each group. (C) Representative images and (D) quantification of PML localization at telomeres in U2OS cells after dimerizing MMS21, MMS21(C215A), Rad52-MMS21/PIAS4 or Rad52-MMS21(C215A)/PIAS4(C342A) for 6 hours. Each dot represents one cell in three independent experiments, more than 55 cells were analyzed in each group. (E) U2OS cells were transfected with HA-SUMO3, 3xHalo-GFP-TRF1 and mCherry-eDHFR-MMS21/MMS21(C215A)/Rad52-MMS21/Rad52-MMS21(C215A). Cells were immunoblotted with anti-TRF2, SUMO1 and β-Actin. (F) Representative images of cells with endogenous PML labeled with Clover, expressing mCh-eDHFR-MMS21/MMS21(C215A) or (H) Rad52-MMS21/MMS21(C215A) and 3xHalo-TRF1 with the addition of the dimerizer at indicated time points. Yellow arrows indicate APBs. (G) Quantification of newly formed APBs per cell after dimerizing MMS21(C215A)/Rad52-MMS21/PIAS4(C342A)/Rad52-PIAS4(C342A) for 2 hours. More than 25 cells were analyzed in each group from three independent experiments. Scale bars, 5 μm.

Journal: bioRxiv

Article Title: Telomeric SUMO level influences the choices of APB formation pathways and ALT efficiency

doi: 10.1101/2025.01.16.633463

Figure Lengend Snippet: (A) Representative images and (B) quantification of SUMO1/2/3 intensity per unit of MMS21/PIAS4 or Rad52-MMS21/PIAS4/MMS21(C215A)/PIAS4(C342A) intensity on telomeres in U2OS cells. Each dot represents one cell in three independent experiments, more than 47 cells were analyzed in each group. (C) Representative images and (D) quantification of PML localization at telomeres in U2OS cells after dimerizing MMS21, MMS21(C215A), Rad52-MMS21/PIAS4 or Rad52-MMS21(C215A)/PIAS4(C342A) for 6 hours. Each dot represents one cell in three independent experiments, more than 55 cells were analyzed in each group. (E) U2OS cells were transfected with HA-SUMO3, 3xHalo-GFP-TRF1 and mCherry-eDHFR-MMS21/MMS21(C215A)/Rad52-MMS21/Rad52-MMS21(C215A). Cells were immunoblotted with anti-TRF2, SUMO1 and β-Actin. (F) Representative images of cells with endogenous PML labeled with Clover, expressing mCh-eDHFR-MMS21/MMS21(C215A) or (H) Rad52-MMS21/MMS21(C215A) and 3xHalo-TRF1 with the addition of the dimerizer at indicated time points. Yellow arrows indicate APBs. (G) Quantification of newly formed APBs per cell after dimerizing MMS21(C215A)/Rad52-MMS21/PIAS4(C342A)/Rad52-PIAS4(C342A) for 2 hours. More than 25 cells were analyzed in each group from three independent experiments. Scale bars, 5 μm.

Techniques: Transfection, Labeling, Expressing

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Structured Review

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1) Product Images from "The Role of SUMO1 Modification of SOX9 in Cartilage Development Stimulated by Zinc Ions in Mice"

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