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dna fe2 staining kit beyotime s10070s 1 × working dapi beyotime c1005  (Beyotime)


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    Beyotime dna fe2 staining kit beyotime s10070s 1 × working dapi beyotime c1005
    Dna Fe2 Staining Kit Beyotime S10070s 1 × Working Dapi Beyotime C1005, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 33153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dapi-stained+dna/DAPI/pm41475687-94-40-44
    Average 99 stars, based on 33153 article reviews
    dna fe2 staining kit beyotime s10070s 1 × working dapi beyotime c1005 - by Bioz Stars, 2026-10
    99/100 stars

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    Virus:


    Recombinant:


    Staining:


    Article Title: Extrachromosomal DNA Couples with ATM-mediated DNA Damage Response for Genome Instability in Tumors
    Article Snippet: Protein concentrations of cell lysates were determined by BCA assay (Beyotime, P0009). .. Then, the cells were washed with PBS, and DNA was stained with DAPI for 5 min at RT, mounted with Antifade Mounting Medium with DAPI (Beyotime, P0131) and viewed under the A1R confocal microscope (Nikon, Japan). .. Proteins were then transferred to PVDF membrane (Millipore, ISEQ00010) and blocked with non-fat milk solution (5%, v/v) for 1 h, then washed three times in 1×TBST for approximately 5 min each.

    Article Title: Light-triggered molecular mechanotherapy of tumor using membrane-mimicking conjugated oligoelectrolytes
    Article Snippet: .. Subsequently, cells were incubated with DAPI staining solution (a membrane-impermeable nuclear stain used to assess cell membrane integrity by binding to DNA in cells with compromised membranes, Beyotime Biotechnology, China) at a concentration recommended by the manufacturer for 15 minutes at room temperature in the dark. ..

    Article Title: ANXA6/TRPV2 axis promotes lymphatic metastasis in head and neck squamous cell carcinoma by inducing autophagy
    Article Snippet: .. We stained nuclear DNA with DAPI (C1006; Beyotime, Shanghai, China) and used a Nikon confocal system for image capture. .. Cell Counting Kit-8 (CCK-8; Beyotime, Shanghai, China) and EdU (RIBOBIO, Guangzhou, China) assays were used to detect the proliferation of HNSCC cells.

    Article Title: GluN2A Mediates PS-Induced Depressive-Like Behavior by Activating CaMKII and Inhibiting Myelinization.
    Article Snippet: After blocking with goat serum, tissue sections were incubated overnight at 4 °C with antibody, washed four times with PBST for 3 min each, and then incubated for 1 h at 37 °C with Cy3-conjugated goat anti-rabbit IgG secondary antibody (1:100; BA1032, BOSTER Biological Technology). .. The sections were then incubated for 5 min with DNA stain 2-(4-amidinophenyl)6-indolecarbamidine dihydrochloride (DAPI, C1002, Beyotime Biotech, Shanghai, China), washed four times with PBST for 3 min each, and mounted in glycerol (50% v/v). .. Fluorescence microscopy was used to view slides (BX53, Olympus, Tokyo, Japan) and the fluorescence intensity measured with Image-Pro Plus (Version 6.0, Media Cybernetics, Rockville, MD, USA).

    Article Title: ANXA6/TRPV2 axis promotes lymphatic metastasis in head and neck squamous cell carcinoma by inducing autophagy.
    Article Snippet: .. We stained nuclear DNA with DAPI (C1006; Beyotime, Shanghai, China) and used a Nikon confocal system for image capture. .. Cell Counting Kit-8 (CCK-8; Beyotime, Shanghai, China) and EdU (RIBOBIO, Guangzhou, China) assays were used to detect the proliferation of HNSCC cells.

    ATPase Assay:


    Bacteria:


    Microscopy:

    Article Title: Extrachromosomal DNA Couples with ATM-mediated DNA Damage Response for Genome Instability in Tumors
    Article Snippet: Protein concentrations of cell lysates were determined by BCA assay (Beyotime, P0009). .. Then, the cells were washed with PBS, and DNA was stained with DAPI for 5 min at RT, mounted with Antifade Mounting Medium with DAPI (Beyotime, P0131) and viewed under the A1R confocal microscope (Nikon, Japan). .. Proteins were then transferred to PVDF membrane (Millipore, ISEQ00010) and blocked with non-fat milk solution (5%, v/v) for 1 h, then washed three times in 1×TBST for approximately 5 min each.

    Incubation:

    Article Title: Light-triggered molecular mechanotherapy of tumor using membrane-mimicking conjugated oligoelectrolytes
    Article Snippet: .. Subsequently, cells were incubated with DAPI staining solution (a membrane-impermeable nuclear stain used to assess cell membrane integrity by binding to DNA in cells with compromised membranes, Beyotime Biotechnology, China) at a concentration recommended by the manufacturer for 15 minutes at room temperature in the dark. ..

    Article Title: GluN2A Mediates PS-Induced Depressive-Like Behavior by Activating CaMKII and Inhibiting Myelinization.
    Article Snippet: After blocking with goat serum, tissue sections were incubated overnight at 4 °C with antibody, washed four times with PBST for 3 min each, and then incubated for 1 h at 37 °C with Cy3-conjugated goat anti-rabbit IgG secondary antibody (1:100; BA1032, BOSTER Biological Technology). .. The sections were then incubated for 5 min with DNA stain 2-(4-amidinophenyl)6-indolecarbamidine dihydrochloride (DAPI, C1002, Beyotime Biotech, Shanghai, China), washed four times with PBST for 3 min each, and mounted in glycerol (50% v/v). .. Fluorescence microscopy was used to view slides (BX53, Olympus, Tokyo, Japan) and the fluorescence intensity measured with Image-Pro Plus (Version 6.0, Media Cybernetics, Rockville, MD, USA).

    Membrane:

    Article Title: Light-triggered molecular mechanotherapy of tumor using membrane-mimicking conjugated oligoelectrolytes
    Article Snippet: .. Subsequently, cells were incubated with DAPI staining solution (a membrane-impermeable nuclear stain used to assess cell membrane integrity by binding to DNA in cells with compromised membranes, Beyotime Biotechnology, China) at a concentration recommended by the manufacturer for 15 minutes at room temperature in the dark. ..


    Binding Assay:

    Article Title: Light-triggered molecular mechanotherapy of tumor using membrane-mimicking conjugated oligoelectrolytes
    Article Snippet: .. Subsequently, cells were incubated with DAPI staining solution (a membrane-impermeable nuclear stain used to assess cell membrane integrity by binding to DNA in cells with compromised membranes, Beyotime Biotechnology, China) at a concentration recommended by the manufacturer for 15 minutes at room temperature in the dark. ..

    Concentration Assay:

    Article Title: Light-triggered molecular mechanotherapy of tumor using membrane-mimicking conjugated oligoelectrolytes
    Article Snippet: .. Subsequently, cells were incubated with DAPI staining solution (a membrane-impermeable nuclear stain used to assess cell membrane integrity by binding to DNA in cells with compromised membranes, Beyotime Biotechnology, China) at a concentration recommended by the manufacturer for 15 minutes at room temperature in the dark. ..

    other:

    Article Title: A dual-action bacteriophage protein Ped1 degrades DNA and blocks replication in Pseudomonas aeruginosa.
    Article Snippet: Reagents and tools table Reagent/resource Reference or source Identifier or catalog number Experimental models List of P. aeruginosa and E. coli strains This paper Dataset EV5 List of Pseudomonas aeruginosa phage PaoP5 This paper Dataset EV5 Recombinant DNA List of plasmids This paper Dataset EV5 Antibodies Mouse anti DDDDK-Tag mAb Abclonal AE005 Mouse anti-His-Tag mAb Abclonal AE003 Mouse anti-GST-Tag mAb Abclonal AE001 HRP-conjugated Goat antiMouse IgG (H+ L) Abclonal AS003 Purified anti-E. coli RNA Polymerase α Antibody BioLegend 663104 Oligonucleotides and other sequence-based reagents oligonucleotide sequence information This paper Dataset EV6 Chemicals, enzymes, and other reagents IPTG (Isopropyl β-D-1thiogalactopyranoside) SigmaAldrich I6758 L-Arabinose SigmaAldrich A3256 Carbenicillin SigmaAldrich C1389 Gentamicin SigmaAldrich G1397 Tetracycline SigmaAldrich T7660 Kanamycin SigmaAldrich K4000 DNase I TransGen Biotech GD201-01 DNase I reaction buffer TransGen Biotech GD201-01 HiTrap Desalting column Cytiva 17546508 HisTrap HP column Cytiva 11000458 GSTrap HP column Cytiva 28401747 Superdex 200 Increase 10/ 300 GL column Cytiva 17546508 10 The EMBO Journal © The Author(s) Reagent/resource Reference or source Identifier or catalog number SYBR Green nucleic acid gel stain Tsingke Biotech N/A One Step TUNEL Apoptosis Assay Kit Beyotime C1086 DAPI (4′,6-diamidino-2phenylindole) SigmaAldrich D9542 PURExpress In Vitro Protein Synthesis Kit New England Biolabs E6800S Murine RNase Inhibitor New England Biolabs M0314 SYBR Green PCR Master Mix abm G891 2×Es Taq MasterMix (Dye) cwbio CW0690S (1 ml) PrimeSTAR Max DNA Polymerase TaKaRa R045Q RNAprep Pure Cell/Bacteria Kit TianGen biotech DP430 Reverse transcriptase kit abm G592 SteadyPure Bacterial Genomic DNA Extraction Kit Accurate Biology AG21007 ECL Plus Kit Cytiva RPN418-1 PVDF membrane Millipore IPVH00010 X-gal (5-Bromo-4-chloro-3indolyl β-Dgalactopyranoside) SigmaAldrich B4252 ONPG (o-Nitrophenyl β-Dgalactopyranoside) SigmaAldrich N1127 QuickMutation Random Mutagenesis Kit Beyotime D0219S PreScission Protease Cytiva 27084301 anti-Flag M2 magnetic beads Sigma M8823 BiodyneTM B Nylon membrane Thermo Scientific 77016 Silver Staining Kit Beyotime P0017S Glutathione beads Beyotime P2258 Software Fiji ImageJ NIH https://imagej.net/ GraphPad Prism GraphPad Software Version 8.0 or later MEGA 7 MEGA software https://www.megasoftware.



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    (A and B) DB008 engages PARP16 and reduces its MARylation in ovarian cancer cells. OVCAR3 cells ectopically expressing FLAG-PARP16 were treated with 0 – 10 µM DB008 for 4 hours. (A) 20 µg of lysate were subjected to click chemistry to visualize DB008 target engagement with in-gel fluorescence (arrow indicates expected band size for FLAG-PARP16). (B) FLAG-PARP16 was immunoprecipitated from the remaining lysate and its MARylation was assessed with Western blotting. (C and D) In situ detection of (C) PARP16 autoMARylation and (D) RPS6 transMARylation. PLA of FLAG and MAR in OVCAR3 cells subjected to Dox-induced expression of FLAG-PARP16 and treated with 0 or 1 µM DB008 for 16 hours (C), or RPS6 and MAR in OVCAR3 cells treated with 0 or 1 µM DB008 for 16 hours (D). <t>DNA</t> was stained with <t>DAPI.</t> Scale bar is 25 µm. (E and F) Quantification of PLA foci in panel C (E) and panel D (F). Each bar represents the mean + SEM; n = 3. Welch’s t-test; ** = p < 0.01. (G) DB008 reduces ribosome MARylation. OVCAR3 cells were treated with 0 or 1 µM DB008 for 16 hours. Ribosomes were fractionated by ultracentrifugation and ribosome MARylation was assessed by Western blotting. RPS6 was used as a loading control. (H and I) DB008 enhances protein synthesis. OVCAR3 cells were treated with 0 – 2 µM DB008 for 16 hours. The rate of protein synthesis was then assessed with a puromycin incorporation assay. (D) Western blot showing puromycin incorporation, COX20, and cleaved caspase-3. β-tubulin was used as a loading control. (E) Quantification of puromycin signal in D. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05, **** = p < 0.0001.
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    (A and B) DB008 engages PARP16 and reduces its MARylation in ovarian cancer cells. OVCAR3 cells ectopically expressing FLAG-PARP16 were treated with 0 – 10 µM DB008 for 4 hours. (A) 20 µg of lysate were subjected to click chemistry to visualize DB008 target engagement with in-gel fluorescence (arrow indicates expected band size for FLAG-PARP16). (B) FLAG-PARP16 was immunoprecipitated from the remaining lysate and its MARylation was assessed with Western blotting. (C and D) In situ detection of (C) PARP16 autoMARylation and (D) RPS6 transMARylation. PLA of FLAG and MAR in OVCAR3 cells subjected to Dox-induced expression of FLAG-PARP16 and treated with 0 or 1 µM DB008 for 16 hours (C), or RPS6 and MAR in OVCAR3 cells treated with 0 or 1 µM DB008 for 16 hours (D). <t>DNA</t> was stained with <t>DAPI.</t> Scale bar is 25 µm. (E and F) Quantification of PLA foci in panel C (E) and panel D (F). Each bar represents the mean + SEM; n = 3. Welch’s t-test; ** = p < 0.01. (G) DB008 reduces ribosome MARylation. OVCAR3 cells were treated with 0 or 1 µM DB008 for 16 hours. Ribosomes were fractionated by ultracentrifugation and ribosome MARylation was assessed by Western blotting. RPS6 was used as a loading control. (H and I) DB008 enhances protein synthesis. OVCAR3 cells were treated with 0 – 2 µM DB008 for 16 hours. The rate of protein synthesis was then assessed with a puromycin incorporation assay. (D) Western blot showing puromycin incorporation, COX20, and cleaved caspase-3. β-tubulin was used as a loading control. (E) Quantification of puromycin signal in D. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05, **** = p < 0.0001.
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    (A and B) DB008 engages PARP16 and reduces its MARylation in ovarian cancer cells. OVCAR3 cells ectopically expressing FLAG-PARP16 were treated with 0 – 10 µM DB008 for 4 hours. (A) 20 µg of lysate were subjected to click chemistry to visualize DB008 target engagement with in-gel fluorescence (arrow indicates expected band size for FLAG-PARP16). (B) FLAG-PARP16 was immunoprecipitated from the remaining lysate and its MARylation was assessed with Western blotting. (C and D) In situ detection of (C) PARP16 autoMARylation and (D) RPS6 transMARylation. PLA of FLAG and MAR in OVCAR3 cells subjected to Dox-induced expression of FLAG-PARP16 and treated with 0 or 1 µM DB008 for 16 hours (C), or RPS6 and MAR in OVCAR3 cells treated with 0 or 1 µM DB008 for 16 hours (D). <t>DNA</t> was stained with <t>DAPI.</t> Scale bar is 25 µm. (E and F) Quantification of PLA foci in panel C (E) and panel D (F). Each bar represents the mean + SEM; n = 3. Welch’s t-test; ** = p < 0.01. (G) DB008 reduces ribosome MARylation. OVCAR3 cells were treated with 0 or 1 µM DB008 for 16 hours. Ribosomes were fractionated by ultracentrifugation and ribosome MARylation was assessed by Western blotting. RPS6 was used as a loading control. (H and I) DB008 enhances protein synthesis. OVCAR3 cells were treated with 0 – 2 µM DB008 for 16 hours. The rate of protein synthesis was then assessed with a puromycin incorporation assay. (D) Western blot showing puromycin incorporation, COX20, and cleaved caspase-3. β-tubulin was used as a loading control. (E) Quantification of puromycin signal in D. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05, **** = p < 0.0001.
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    Beyotime dna fe2 staining kit beyotime s10070s 1 × working dapi beyotime c1005
    (A and B) DB008 engages PARP16 and reduces its MARylation in ovarian cancer cells. OVCAR3 cells ectopically expressing FLAG-PARP16 were treated with 0 – 10 µM DB008 for 4 hours. (A) 20 µg of lysate were subjected to click chemistry to visualize DB008 target engagement with in-gel fluorescence (arrow indicates expected band size for FLAG-PARP16). (B) FLAG-PARP16 was immunoprecipitated from the remaining lysate and its MARylation was assessed with Western blotting. (C and D) In situ detection of (C) PARP16 autoMARylation and (D) RPS6 transMARylation. PLA of FLAG and MAR in OVCAR3 cells subjected to Dox-induced expression of FLAG-PARP16 and treated with 0 or 1 µM DB008 for 16 hours (C), or RPS6 and MAR in OVCAR3 cells treated with 0 or 1 µM DB008 for 16 hours (D). <t>DNA</t> was stained with <t>DAPI.</t> Scale bar is 25 µm. (E and F) Quantification of PLA foci in panel C (E) and panel D (F). Each bar represents the mean + SEM; n = 3. Welch’s t-test; ** = p < 0.01. (G) DB008 reduces ribosome MARylation. OVCAR3 cells were treated with 0 or 1 µM DB008 for 16 hours. Ribosomes were fractionated by ultracentrifugation and ribosome MARylation was assessed by Western blotting. RPS6 was used as a loading control. (H and I) DB008 enhances protein synthesis. OVCAR3 cells were treated with 0 – 2 µM DB008 for 16 hours. The rate of protein synthesis was then assessed with a puromycin incorporation assay. (D) Western blot showing puromycin incorporation, COX20, and cleaved caspase-3. β-tubulin was used as a loading control. (E) Quantification of puromycin signal in D. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05, **** = p < 0.0001.
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    (A and B) DB008 engages PARP16 and reduces its MARylation in ovarian cancer cells. OVCAR3 cells ectopically expressing FLAG-PARP16 were treated with 0 – 10 µM DB008 for 4 hours. (A) 20 µg of lysate were subjected to click chemistry to visualize DB008 target engagement with in-gel fluorescence (arrow indicates expected band size for FLAG-PARP16). (B) FLAG-PARP16 was immunoprecipitated from the remaining lysate and its MARylation was assessed with Western blotting. (C and D) In situ detection of (C) PARP16 autoMARylation and (D) RPS6 transMARylation. PLA of FLAG and MAR in OVCAR3 cells subjected to Dox-induced expression of FLAG-PARP16 and treated with 0 or 1 µM DB008 for 16 hours (C), or RPS6 and MAR in OVCAR3 cells treated with 0 or 1 µM DB008 for 16 hours (D). DNA was stained with DAPI. Scale bar is 25 µm. (E and F) Quantification of PLA foci in panel C (E) and panel D (F). Each bar represents the mean + SEM; n = 3. Welch’s t-test; ** = p < 0.01. (G) DB008 reduces ribosome MARylation. OVCAR3 cells were treated with 0 or 1 µM DB008 for 16 hours. Ribosomes were fractionated by ultracentrifugation and ribosome MARylation was assessed by Western blotting. RPS6 was used as a loading control. (H and I) DB008 enhances protein synthesis. OVCAR3 cells were treated with 0 – 2 µM DB008 for 16 hours. The rate of protein synthesis was then assessed with a puromycin incorporation assay. (D) Western blot showing puromycin incorporation, COX20, and cleaved caspase-3. β-tubulin was used as a loading control. (E) Quantification of puromycin signal in D. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05, **** = p < 0.0001.

    Journal: bioRxiv

    Article Title: PARP16 is a Druggable Regulator of Ribosome MARylation and Protein Homeostasis in Ovarian Cancer Cells

    doi: 10.64898/2026.04.01.715986

    Figure Lengend Snippet: (A and B) DB008 engages PARP16 and reduces its MARylation in ovarian cancer cells. OVCAR3 cells ectopically expressing FLAG-PARP16 were treated with 0 – 10 µM DB008 for 4 hours. (A) 20 µg of lysate were subjected to click chemistry to visualize DB008 target engagement with in-gel fluorescence (arrow indicates expected band size for FLAG-PARP16). (B) FLAG-PARP16 was immunoprecipitated from the remaining lysate and its MARylation was assessed with Western blotting. (C and D) In situ detection of (C) PARP16 autoMARylation and (D) RPS6 transMARylation. PLA of FLAG and MAR in OVCAR3 cells subjected to Dox-induced expression of FLAG-PARP16 and treated with 0 or 1 µM DB008 for 16 hours (C), or RPS6 and MAR in OVCAR3 cells treated with 0 or 1 µM DB008 for 16 hours (D). DNA was stained with DAPI. Scale bar is 25 µm. (E and F) Quantification of PLA foci in panel C (E) and panel D (F). Each bar represents the mean + SEM; n = 3. Welch’s t-test; ** = p < 0.01. (G) DB008 reduces ribosome MARylation. OVCAR3 cells were treated with 0 or 1 µM DB008 for 16 hours. Ribosomes were fractionated by ultracentrifugation and ribosome MARylation was assessed by Western blotting. RPS6 was used as a loading control. (H and I) DB008 enhances protein synthesis. OVCAR3 cells were treated with 0 – 2 µM DB008 for 16 hours. The rate of protein synthesis was then assessed with a puromycin incorporation assay. (D) Western blot showing puromycin incorporation, COX20, and cleaved caspase-3. β-tubulin was used as a loading control. (E) Quantification of puromycin signal in D. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05, **** = p < 0.0001.

    Article Snippet: The stained cells were mounted on microslides using VectaShield Antifade Mounting Medium (Vector laboratories, H-1200-10) with DAPI DNA stain.

    Techniques: Expressing, Drug discovery, Fluorescence, Immunoprecipitation, Western Blot, In Situ, Staining, Control

    (A) Western blot showing PARP16 protein expression is abolished in OVCAR3 cells with PARP16 KO. β-actin was used as a loading control. (B) Immunofluorescence with the MAR detection reagent in OVCAR3 Control or PARP16 KO cells treated with 0 or 1 µM DB008 for 16 hours. The DNA was stained with DAPI. Scale bar = 25 µm. (C) The DB008-dependent increase in protein synthesis requires PARP16 expression. OVCAR3 Control or PARP16 KO cells were treated with 0 or 1 µM DB008 for 16 hours. Protein synthesis was then assessed with a puromycin incorporation assay. β-tubulin was used as a loading control. (D) The DB008-dependent increase in protein aggregation requires PARP16 expression. OVCAR3 Control or PARP16 KO cells were treated with 0 or 1 µM DB008 for 16 hours. Protein aggregation was then assessed with the proteostat aggresome detection reagent. The DNA was stained with DAPI. Scale bar = 25 µm. (E) DB008 reduces growth of control but not OVCAR3 PARP16 KO cells. Cells were treated with 0 or 1 µM DB008 for 1 week. Cell growth was then determined with crystal violet staining. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; ** = p < 0.005; ns indicates no significance. (F) DB008 inhibits the anchorage-independent growth of ovarian cancer cells. OVCAR3 cells were grown in soft agar with media containing 0 – 1 µM DB008 for 3 weeks, replenishing media once per week. Anchorage-independent growth was then measured with crystal violet staining. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05, **** = p < 0.0001.

    Journal: bioRxiv

    Article Title: PARP16 is a Druggable Regulator of Ribosome MARylation and Protein Homeostasis in Ovarian Cancer Cells

    doi: 10.64898/2026.04.01.715986

    Figure Lengend Snippet: (A) Western blot showing PARP16 protein expression is abolished in OVCAR3 cells with PARP16 KO. β-actin was used as a loading control. (B) Immunofluorescence with the MAR detection reagent in OVCAR3 Control or PARP16 KO cells treated with 0 or 1 µM DB008 for 16 hours. The DNA was stained with DAPI. Scale bar = 25 µm. (C) The DB008-dependent increase in protein synthesis requires PARP16 expression. OVCAR3 Control or PARP16 KO cells were treated with 0 or 1 µM DB008 for 16 hours. Protein synthesis was then assessed with a puromycin incorporation assay. β-tubulin was used as a loading control. (D) The DB008-dependent increase in protein aggregation requires PARP16 expression. OVCAR3 Control or PARP16 KO cells were treated with 0 or 1 µM DB008 for 16 hours. Protein aggregation was then assessed with the proteostat aggresome detection reagent. The DNA was stained with DAPI. Scale bar = 25 µm. (E) DB008 reduces growth of control but not OVCAR3 PARP16 KO cells. Cells were treated with 0 or 1 µM DB008 for 1 week. Cell growth was then determined with crystal violet staining. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; ** = p < 0.005; ns indicates no significance. (F) DB008 inhibits the anchorage-independent growth of ovarian cancer cells. OVCAR3 cells were grown in soft agar with media containing 0 – 1 µM DB008 for 3 weeks, replenishing media once per week. Anchorage-independent growth was then measured with crystal violet staining. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05, **** = p < 0.0001.

    Article Snippet: The stained cells were mounted on microslides using VectaShield Antifade Mounting Medium (Vector laboratories, H-1200-10) with DAPI DNA stain.

    Techniques: Western Blot, Expressing, Control, Immunofluorescence, Staining

    (A and B) Inhibition by DB008 requires PARP16 C169. (A) IP-Western showing PARP16 MARylation ( top panel ) or PARylation levels in whole cell extracts (WCE; bottom panel ) of OVCAR3 PARP16 KO cells stably expressing FLAG-PARP16 or FLAG-PARP16 C169S after treatment with 1 µM DB008 for 4 hours. (B) Quantification of MAR signal from A. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05; ns indicates no significance. (C) Anchorage-independent growth is unaffected in cells expressing PARP16 C169S. OVCAR3 PARP16 KO cells stably expressing PARP16 WT or PARP16 C169S were grown in soft agar with media containing 0 or 1 µM DB008 for 3 weeks, replenishing media once per week. Anchorage-independent growth was then measured with crystal violet staining. Quantification of crystal violet absorbance in B. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05; ns indicates no significance. (D and E) The DB008-dependent increase in protein synthesis requires PARP16 C169. (D) OVCAR3 PARP16 KO cells stably expressing PARP16 WT or PARP16 C169S were treated with 0 or 1 µM DB008 for 16 hours. Protein synthesis was then assessed with a puromycin incorporation assay. β-tubulin was used as a loading control. (E) Quantification of puromycin signal in D. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; ** = p < 0.01; ns indicates no significance. (F) The DB008-dependent increase in protein aggregation requires PARP16 C169. OVCAR3 PARP16 KO cells stably expressing PARP16 WT or PARP16 C169S were treated with 0 or 1 µM DB008 for 16 hours. Protein aggregation was then assessed with the proteostat aggresome detection reagent. The DNA was stained with DAPI. Scale bar = 25 µm.

    Journal: bioRxiv

    Article Title: PARP16 is a Druggable Regulator of Ribosome MARylation and Protein Homeostasis in Ovarian Cancer Cells

    doi: 10.64898/2026.04.01.715986

    Figure Lengend Snippet: (A and B) Inhibition by DB008 requires PARP16 C169. (A) IP-Western showing PARP16 MARylation ( top panel ) or PARylation levels in whole cell extracts (WCE; bottom panel ) of OVCAR3 PARP16 KO cells stably expressing FLAG-PARP16 or FLAG-PARP16 C169S after treatment with 1 µM DB008 for 4 hours. (B) Quantification of MAR signal from A. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05; ns indicates no significance. (C) Anchorage-independent growth is unaffected in cells expressing PARP16 C169S. OVCAR3 PARP16 KO cells stably expressing PARP16 WT or PARP16 C169S were grown in soft agar with media containing 0 or 1 µM DB008 for 3 weeks, replenishing media once per week. Anchorage-independent growth was then measured with crystal violet staining. Quantification of crystal violet absorbance in B. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; * = p < 0.05; ns indicates no significance. (D and E) The DB008-dependent increase in protein synthesis requires PARP16 C169. (D) OVCAR3 PARP16 KO cells stably expressing PARP16 WT or PARP16 C169S were treated with 0 or 1 µM DB008 for 16 hours. Protein synthesis was then assessed with a puromycin incorporation assay. β-tubulin was used as a loading control. (E) Quantification of puromycin signal in D. Each bar represents the mean + SEM; n = 3. Bars marked with asterisks are significantly different from the control; ANOVA; ** = p < 0.01; ns indicates no significance. (F) The DB008-dependent increase in protein aggregation requires PARP16 C169. OVCAR3 PARP16 KO cells stably expressing PARP16 WT or PARP16 C169S were treated with 0 or 1 µM DB008 for 16 hours. Protein aggregation was then assessed with the proteostat aggresome detection reagent. The DNA was stained with DAPI. Scale bar = 25 µm.

    Article Snippet: The stained cells were mounted on microslides using VectaShield Antifade Mounting Medium (Vector laboratories, H-1200-10) with DAPI DNA stain.

    Techniques: Inhibition, Western Blot, Stable Transfection, Expressing, Control, Staining