anti proliferating cell nuclear antigen pcna  (BioLegend)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 91

    Structured Review

    BioLegend anti proliferating cell nuclear antigen pcna
    Anti Proliferating Cell Nuclear Antigen Pcna, supplied by BioLegend, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti proliferating cell nuclear antigen pcna/product/BioLegend
    Average 91 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    anti proliferating cell nuclear antigen pcna - by Bioz Stars, 2020-07
    91/100 stars

    Images

    Related Articles

    Marker:

    Article Title: G-CSF and G-CSFR are highly expressed in human gastric and colon cancers and promote carcinoma cell proliferation and migration
    Article Snippet: .. Anti-proliferating cell nuclear antigen (PCNA) (clone PCNA01, Biolegend) was used as a proliferation marker. .. Cancer stem-like populations in carcinoma cell lines were examined by staining with Aldefluor (Stem Cell Technologies, Vancouver, BC, Canada) and anti-CD44-APC (Biolegend clone BJ18) using the manufacturer's Aldefluor staining protocol compared with isotype control.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    BioLegend anti pcna
    Proliferation of HSCs was reduced by deficiency of NOX1 and NOX4 in response to liver injury and PDGF. (A) HSC proliferation is reduced in NOX1KO and NOX4KO mice after CCl 4 injury. HSC proliferation in liver was presented by <t>PCNA</t> immunofluorescence microscopy. <t>Desmin</t> is a specific marker of HSCs (Red). PCNA is a marker of proliferation (Green). Original magnification X10. (B) Proliferation was reduced in HSCs lacking NOX1 and NOX4 in response to PDGF (10 ng/ml) for 24 h. Proliferative HSCs were presented by Ki67 (Red) immunofluorescence microscopy. Nuclei were presented by DAPI (Blue). Original magnification X10. (C) Graph of PCNA-positive HSCs in vivo. (D) Graph of Ki67-positive HSCs in vitro.
    Anti Pcna, supplied by BioLegend, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti pcna/product/BioLegend
    Average 92 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    anti pcna - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    85
    BioLegend fitc conjugated anti pcna antibody
    PDGF-mediated inhibition of p27Kip1 by miR-221 Promotes Cell proliferation. (A, B): AsPC-1 cells were transfected with a negative control mimic, the miR-221 mimic, antisense oligonucleotides to miR-221 (anti-mIR-221) or p27Kip1 siRNA. The cells were then treated with PDGF-BB (20 ng/ml) for 24 h and subjected to qRT-PCR (A) and Western blot analysis (B) for p27Kip1. (C): AsPC-1 cells were transfected with negative control or p27Kip1 siRNA and subjected to the MTT cell proliferation assay (The results are indicated as the absorbance readings at 490 nm). (C) (D, E): AsPC-1 cells were transfected with a negative control mimic, the miR-221 mimic, antisense oligonucleotides to miR-221 (anti-mIR-221) or p27Kip1 siRNA. The cells were then treated with PDGF-BB (20 ng/ml) for 24 h and subjected to staining with a <t>FITC-conjugated</t> antibody against the proliferation marker <t>PCNA</t> and DAPI (E). In total, 150 cells were counted per condition, and the percentage of PCNA-positive cells is presented (D). All experiments in this figure were carried out in triplicate, and the results are displayed as the means ± SD.
    Fitc Conjugated Anti Pcna Antibody, supplied by BioLegend, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fitc conjugated anti pcna antibody/product/BioLegend
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    fitc conjugated anti pcna antibody - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    94
    BioLegend pe pcna
    The assessment of proliferation-related biomarkers in breast cancer patient-derived CTCs. a Ki67 staining in PanCK+ and <t>CD44+/CD24−</t> CTC subtypes isolated from breast cancer patient blood. Scale bar = 10 μm. b uPAR and int-β1 expression in single CTCs designated as Ki67 Low ( n = 60) or Ki67 High ( n = 48). p -value was calculated by two-way matched ANOVA. Despite a statistically significant overall difference, 20–30% individual CTCs in both groups had similar uPAR and int-β1 expression. c Ki67 expression in single CTCs designated as uPAR−/int-β1− ( n = 17) or uPAR+/int-β1+ ( n = 11). p -value calculated by unpaired t -test. Of note, 7 out of 17 uPAR/int-β1 CTCs had higher Ki67 expression than the rest of the group. d Demonstration of Ki67, uPAR, and int-β1 staining of single CTCs by DEPArray TM . Note that the CTC in the third row, is designated as uPAR−/int-β1, but the Ki67 expression is higher than the Ki67 Low CTC shown in the first row . e Flowchart depicting PBMC processing for concomitant cell surface and nuclear staining for single-step analysis by flow cytometry. <t>PCNA</t> was replaced by cleaved PARP (Asp214) in the last eight samples to accommodate an apoptosis marker in the nine-color panel . f MTN index of Ki67 Low vs. Ki67 High CTCs assessed by the Permiflow TM method. g Difference in Ki67 status of BCBM vs. no BCBM patient-derived CTCs ( n = 8). p- value calculated by two-way matched ANOVA test. h Difference in uPAR/int-β1 expression status in BCBM vs. no BCBM patient-derived CTCs ( n = 8). p- value calculated by two-way matched ANOVA test. i uPAR and int-β1 expression in CTC groups designated as Ki67 Low or Ki67 High ( n = 32). p -value calculated by two-way matched ANOVA. j Ki67 expression in CTC groups designated as uPAR−/int-β1− or uPAR+/int-β1+ ( n = 16). p -value calculated by unpaired t -test. k Circos plot depicting cancer-related gene exon insertions/deletion (in/del) mutations in uPAR+/int-β1+ and uPAR−/int-β1− CTC subsets obtained from BCBM and no BCBM patients. From outer to inner circle : uPAR+/int-β1+ and uPAR−/int β1− CTCs derived from BCBM patient; uPAR+/int β1+ and uPAR−/int-β1− CTCs derived from no BCBM patient. 881/884 in/del mutations found in BCBM subsets, whereas 434/211 in/del in no BCBM subsets. Error bars represent s.e.m
    Pe Pcna, supplied by BioLegend, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pe pcna/product/BioLegend
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pe pcna - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    91
    BioLegend anti pcna monoclonal antibody
    PARI localization at replication forks and DNA damage sites. (A) HEK 293T (top) and NIH 3T3 (bottom) cells transiently transfected with <t>pCAG-FLAG-PARI</t> and pCAG-PARI-6×His plasmids were immunostained with anti-FLAG and anti-6×His antibodies, respectively. DAPI was used to visualize the nuclei. Scale bars, 5 μm. (B) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI immunostained for FLAG (red) and <t>PCNA</t> (green). Representative images of early, mid-, and late S phase cells are presented. The insets show higher-magnification views of the boxed regions. Scale bars, 5 μm. (C) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI and pulse-labeled with BrdU were stained with anti-FLAG (red) and anti-BrdU (green) antibodies. (D) iPOND analysis of 293T cells transfected with pCAG-FLAG-PARI and pulse-labeled with EdU. Nascent replication forks were captured by the click reaction, and FLAG-PARI and PCNA were detected by Western blotting (WB). The input lanes represent 2.4% of the cell lysates used for the iPOND elution. (E) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI were treated with MMC for 24 h and immunostained for FLAG (green) and PCNA (red). (F) NIH 3T3 cells transiently transfected with plasmids that express FLAG-tagged full-length PARI (top) or PIP box deletion mutant PARIΔPIP (bottom) constructs were stained with anti-FLAG antibody (green). Nuclei were counterstained with DAPI (blue). (G) U2OS cells transfected with a plasmid that expresses GFP-human PARI were laser irradiated (between arrowheads) during S phase (left) or G 1 /G 2 phase (right), and live-cell imaging was carried out. Representative images at the indicated time points are shown. Scale bars, 5 μm.
    Anti Pcna Monoclonal Antibody, supplied by BioLegend, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti pcna monoclonal antibody/product/BioLegend
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti pcna monoclonal antibody - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    Image Search Results


    Proliferation of HSCs was reduced by deficiency of NOX1 and NOX4 in response to liver injury and PDGF. (A) HSC proliferation is reduced in NOX1KO and NOX4KO mice after CCl 4 injury. HSC proliferation in liver was presented by PCNA immunofluorescence microscopy. Desmin is a specific marker of HSCs (Red). PCNA is a marker of proliferation (Green). Original magnification X10. (B) Proliferation was reduced in HSCs lacking NOX1 and NOX4 in response to PDGF (10 ng/ml) for 24 h. Proliferative HSCs were presented by Ki67 (Red) immunofluorescence microscopy. Nuclei were presented by DAPI (Blue). Original magnification X10. (C) Graph of PCNA-positive HSCs in vivo. (D) Graph of Ki67-positive HSCs in vitro.

    Journal: PLoS ONE

    Article Title: Deficiency of NOX1 or NOX4 Prevents Liver Inflammation and Fibrosis in Mice through Inhibition of Hepatic Stellate Cell Activation

    doi: 10.1371/journal.pone.0129743

    Figure Lengend Snippet: Proliferation of HSCs was reduced by deficiency of NOX1 and NOX4 in response to liver injury and PDGF. (A) HSC proliferation is reduced in NOX1KO and NOX4KO mice after CCl 4 injury. HSC proliferation in liver was presented by PCNA immunofluorescence microscopy. Desmin is a specific marker of HSCs (Red). PCNA is a marker of proliferation (Green). Original magnification X10. (B) Proliferation was reduced in HSCs lacking NOX1 and NOX4 in response to PDGF (10 ng/ml) for 24 h. Proliferative HSCs were presented by Ki67 (Red) immunofluorescence microscopy. Nuclei were presented by DAPI (Blue). Original magnification X10. (C) Graph of PCNA-positive HSCs in vivo. (D) Graph of Ki67-positive HSCs in vitro.

    Article Snippet: For immunofluorescent staining, the sections were incubated with anti-HNE (1:200, Alpha Diagnostics, San Antonio, TX), anti-PCNA (1:200, Biolegend Inc., San Diego, CA), anti-desmin (1:200, Thermo Fisher Scientific, Fremont, CA), or anti-Ki67 (1:200, GeneTEX, INC. Irvine, CA) antibodies and Alexa Fluor 594- and Alexa Fluor 488-conjugated secondary antibodies (1:200, Invitrogen, Carlsbad, CA) and imaged with fluorescent microscopy.

    Techniques: Mouse Assay, Immunofluorescence, Microscopy, Marker, In Vivo, In Vitro

    PDGF-mediated inhibition of p27Kip1 by miR-221 Promotes Cell proliferation. (A, B): AsPC-1 cells were transfected with a negative control mimic, the miR-221 mimic, antisense oligonucleotides to miR-221 (anti-mIR-221) or p27Kip1 siRNA. The cells were then treated with PDGF-BB (20 ng/ml) for 24 h and subjected to qRT-PCR (A) and Western blot analysis (B) for p27Kip1. (C): AsPC-1 cells were transfected with negative control or p27Kip1 siRNA and subjected to the MTT cell proliferation assay (The results are indicated as the absorbance readings at 490 nm). (C) (D, E): AsPC-1 cells were transfected with a negative control mimic, the miR-221 mimic, antisense oligonucleotides to miR-221 (anti-mIR-221) or p27Kip1 siRNA. The cells were then treated with PDGF-BB (20 ng/ml) for 24 h and subjected to staining with a FITC-conjugated antibody against the proliferation marker PCNA and DAPI (E). In total, 150 cells were counted per condition, and the percentage of PCNA-positive cells is presented (D). All experiments in this figure were carried out in triplicate, and the results are displayed as the means ± SD.

    Journal: PLoS ONE

    Article Title: MicroRNA-221 Mediates the Effects of PDGF-BB on Migration, Proliferation, and the Epithelial-Mesenchymal Transition in Pancreatic Cancer Cells

    doi: 10.1371/journal.pone.0071309

    Figure Lengend Snippet: PDGF-mediated inhibition of p27Kip1 by miR-221 Promotes Cell proliferation. (A, B): AsPC-1 cells were transfected with a negative control mimic, the miR-221 mimic, antisense oligonucleotides to miR-221 (anti-mIR-221) or p27Kip1 siRNA. The cells were then treated with PDGF-BB (20 ng/ml) for 24 h and subjected to qRT-PCR (A) and Western blot analysis (B) for p27Kip1. (C): AsPC-1 cells were transfected with negative control or p27Kip1 siRNA and subjected to the MTT cell proliferation assay (The results are indicated as the absorbance readings at 490 nm). (C) (D, E): AsPC-1 cells were transfected with a negative control mimic, the miR-221 mimic, antisense oligonucleotides to miR-221 (anti-mIR-221) or p27Kip1 siRNA. The cells were then treated with PDGF-BB (20 ng/ml) for 24 h and subjected to staining with a FITC-conjugated antibody against the proliferation marker PCNA and DAPI (E). In total, 150 cells were counted per condition, and the percentage of PCNA-positive cells is presented (D). All experiments in this figure were carried out in triplicate, and the results are displayed as the means ± SD.

    Article Snippet: For the proliferating cell nuclear antigen (PCNA) staining , AsPC-1 cells were stained with a FITC-conjugated anti-PCNA antibody (clone PC10) from Biolegend as well as DAPI.

    Techniques: Inhibition, Transfection, Negative Control, Quantitative RT-PCR, Western Blot, MTT Assay, Proliferation Assay, Staining, Marker

    PDGF promoted cell migration and proliferation and altered the EMT phenotype in AsPC-1 cells. AsPC-1 cells incubated with vehicle or PDGF-BB (20 ng/ml) were subjected to the scratch wound assay (A) and Matrigel transmembrane invasion assay (B). The results are the mean±SD. of triplicate measurements of three independent experiments. AsPC-1 cells incubated with vehicle or PDGF-BB (20 ng/ml) for 24 h were subjected to the MTT cell proliferation assay (The results are indicated as the absorbance readings at 490 nm) (C) or stained with a FITC-conjugated antibody against the proliferation marker PCNA and DAPI (150 cells were counted per condition, and the percentage of PCNA-positive cells is presented.) (D). The results are the mean±SD. for triplicate assays of three independent experiments. Real time-PCR was used to determine the mRNA levels to evaluate the upregulated expression of transcription factors (E) and EMT-specific genes (F) in AsPC-1 cells incubated with vehicle or PDGF-BB (20 ng/ml, 24 hr). The relative mRNA levels were normalized to GAPDH. All of the treatments in this figure were carried out in triplicate, and the results are displayed as the means ± SD. G: Photomicrographs of AsPC-1 cells incubated with vehicle or PDGF-BB (20 ng/ml, 24 hr). Original magnification, 200X.

    Journal: PLoS ONE

    Article Title: MicroRNA-221 Mediates the Effects of PDGF-BB on Migration, Proliferation, and the Epithelial-Mesenchymal Transition in Pancreatic Cancer Cells

    doi: 10.1371/journal.pone.0071309

    Figure Lengend Snippet: PDGF promoted cell migration and proliferation and altered the EMT phenotype in AsPC-1 cells. AsPC-1 cells incubated with vehicle or PDGF-BB (20 ng/ml) were subjected to the scratch wound assay (A) and Matrigel transmembrane invasion assay (B). The results are the mean±SD. of triplicate measurements of three independent experiments. AsPC-1 cells incubated with vehicle or PDGF-BB (20 ng/ml) for 24 h were subjected to the MTT cell proliferation assay (The results are indicated as the absorbance readings at 490 nm) (C) or stained with a FITC-conjugated antibody against the proliferation marker PCNA and DAPI (150 cells were counted per condition, and the percentage of PCNA-positive cells is presented.) (D). The results are the mean±SD. for triplicate assays of three independent experiments. Real time-PCR was used to determine the mRNA levels to evaluate the upregulated expression of transcription factors (E) and EMT-specific genes (F) in AsPC-1 cells incubated with vehicle or PDGF-BB (20 ng/ml, 24 hr). The relative mRNA levels were normalized to GAPDH. All of the treatments in this figure were carried out in triplicate, and the results are displayed as the means ± SD. G: Photomicrographs of AsPC-1 cells incubated with vehicle or PDGF-BB (20 ng/ml, 24 hr). Original magnification, 200X.

    Article Snippet: For the proliferating cell nuclear antigen (PCNA) staining , AsPC-1 cells were stained with a FITC-conjugated anti-PCNA antibody (clone PC10) from Biolegend as well as DAPI.

    Techniques: Migration, Incubation, Scratch Wound Assay Assay, Invasion Assay, MTT Assay, Proliferation Assay, Staining, Marker, Real-time Polymerase Chain Reaction, Expressing

    miR-221 is critical for the PDGF-mediated epithelial-mesenchymal transition phenotype, migration, and growth of AsPC-1 cells. AsPC-1 cells were transfected with a negative control, the miR-221 mimic, or antisense oligonucleotides to miR-221 (anti-mIR-221). The cells were then treated with PDGF-BB (20 ng/ml) for 24 h and subjected to qRT-PCR (A, C) or Western blot analysis (B) of the transcription factors and EMT-specific gene markers. AsPC-1 cells were transfected with a negative control, the miR-221 mimic, or antisense oligonucleotides to miR-221 (anti-mIR-221) and subjected to the Matrigel transmembrane invasion assay (D) in the presence or absence of 20 ng/ml PDGF-BB. AsPC-1 cells were transfected with a negative control, the miR-221 mimic, or antisense oligonucleotides to miR-221 (anti-mIR-221), followed by PDGF-BB treatment for 24 h, and then were stained with a FITC-conjugated antibody against the proliferation marker PCNA and DAPI. In total, 150 cells were counted per condition, and the percentage of PCNA-positive cells (E) is presented. All treatment experiments in this figure were carried out in triplicate, and the results are displayed as the means ± SD.

    Journal: PLoS ONE

    Article Title: MicroRNA-221 Mediates the Effects of PDGF-BB on Migration, Proliferation, and the Epithelial-Mesenchymal Transition in Pancreatic Cancer Cells

    doi: 10.1371/journal.pone.0071309

    Figure Lengend Snippet: miR-221 is critical for the PDGF-mediated epithelial-mesenchymal transition phenotype, migration, and growth of AsPC-1 cells. AsPC-1 cells were transfected with a negative control, the miR-221 mimic, or antisense oligonucleotides to miR-221 (anti-mIR-221). The cells were then treated with PDGF-BB (20 ng/ml) for 24 h and subjected to qRT-PCR (A, C) or Western blot analysis (B) of the transcription factors and EMT-specific gene markers. AsPC-1 cells were transfected with a negative control, the miR-221 mimic, or antisense oligonucleotides to miR-221 (anti-mIR-221) and subjected to the Matrigel transmembrane invasion assay (D) in the presence or absence of 20 ng/ml PDGF-BB. AsPC-1 cells were transfected with a negative control, the miR-221 mimic, or antisense oligonucleotides to miR-221 (anti-mIR-221), followed by PDGF-BB treatment for 24 h, and then were stained with a FITC-conjugated antibody against the proliferation marker PCNA and DAPI. In total, 150 cells were counted per condition, and the percentage of PCNA-positive cells (E) is presented. All treatment experiments in this figure were carried out in triplicate, and the results are displayed as the means ± SD.

    Article Snippet: For the proliferating cell nuclear antigen (PCNA) staining , AsPC-1 cells were stained with a FITC-conjugated anti-PCNA antibody (clone PC10) from Biolegend as well as DAPI.

    Techniques: Migration, Transfection, Negative Control, Quantitative RT-PCR, Western Blot, Invasion Assay, Staining, Marker

    The assessment of proliferation-related biomarkers in breast cancer patient-derived CTCs. a Ki67 staining in PanCK+ and CD44+/CD24− CTC subtypes isolated from breast cancer patient blood. Scale bar = 10 μm. b uPAR and int-β1 expression in single CTCs designated as Ki67 Low ( n = 60) or Ki67 High ( n = 48). p -value was calculated by two-way matched ANOVA. Despite a statistically significant overall difference, 20–30% individual CTCs in both groups had similar uPAR and int-β1 expression. c Ki67 expression in single CTCs designated as uPAR−/int-β1− ( n = 17) or uPAR+/int-β1+ ( n = 11). p -value calculated by unpaired t -test. Of note, 7 out of 17 uPAR/int-β1 CTCs had higher Ki67 expression than the rest of the group. d Demonstration of Ki67, uPAR, and int-β1 staining of single CTCs by DEPArray TM . Note that the CTC in the third row, is designated as uPAR−/int-β1, but the Ki67 expression is higher than the Ki67 Low CTC shown in the first row . e Flowchart depicting PBMC processing for concomitant cell surface and nuclear staining for single-step analysis by flow cytometry. PCNA was replaced by cleaved PARP (Asp214) in the last eight samples to accommodate an apoptosis marker in the nine-color panel . f MTN index of Ki67 Low vs. Ki67 High CTCs assessed by the Permiflow TM method. g Difference in Ki67 status of BCBM vs. no BCBM patient-derived CTCs ( n = 8). p- value calculated by two-way matched ANOVA test. h Difference in uPAR/int-β1 expression status in BCBM vs. no BCBM patient-derived CTCs ( n = 8). p- value calculated by two-way matched ANOVA test. i uPAR and int-β1 expression in CTC groups designated as Ki67 Low or Ki67 High ( n = 32). p -value calculated by two-way matched ANOVA. j Ki67 expression in CTC groups designated as uPAR−/int-β1− or uPAR+/int-β1+ ( n = 16). p -value calculated by unpaired t -test. k Circos plot depicting cancer-related gene exon insertions/deletion (in/del) mutations in uPAR+/int-β1+ and uPAR−/int-β1− CTC subsets obtained from BCBM and no BCBM patients. From outer to inner circle : uPAR+/int-β1+ and uPAR−/int β1− CTCs derived from BCBM patient; uPAR+/int β1+ and uPAR−/int-β1− CTCs derived from no BCBM patient. 881/884 in/del mutations found in BCBM subsets, whereas 434/211 in/del in no BCBM subsets. Error bars represent s.e.m

    Journal: Nature Communications

    Article Title: Molecular characterization of breast cancer CTCs associated with brain metastasis

    doi: 10.1038/s41467-017-00196-1

    Figure Lengend Snippet: The assessment of proliferation-related biomarkers in breast cancer patient-derived CTCs. a Ki67 staining in PanCK+ and CD44+/CD24− CTC subtypes isolated from breast cancer patient blood. Scale bar = 10 μm. b uPAR and int-β1 expression in single CTCs designated as Ki67 Low ( n = 60) or Ki67 High ( n = 48). p -value was calculated by two-way matched ANOVA. Despite a statistically significant overall difference, 20–30% individual CTCs in both groups had similar uPAR and int-β1 expression. c Ki67 expression in single CTCs designated as uPAR−/int-β1− ( n = 17) or uPAR+/int-β1+ ( n = 11). p -value calculated by unpaired t -test. Of note, 7 out of 17 uPAR/int-β1 CTCs had higher Ki67 expression than the rest of the group. d Demonstration of Ki67, uPAR, and int-β1 staining of single CTCs by DEPArray TM . Note that the CTC in the third row, is designated as uPAR−/int-β1, but the Ki67 expression is higher than the Ki67 Low CTC shown in the first row . e Flowchart depicting PBMC processing for concomitant cell surface and nuclear staining for single-step analysis by flow cytometry. PCNA was replaced by cleaved PARP (Asp214) in the last eight samples to accommodate an apoptosis marker in the nine-color panel . f MTN index of Ki67 Low vs. Ki67 High CTCs assessed by the Permiflow TM method. g Difference in Ki67 status of BCBM vs. no BCBM patient-derived CTCs ( n = 8). p- value calculated by two-way matched ANOVA test. h Difference in uPAR/int-β1 expression status in BCBM vs. no BCBM patient-derived CTCs ( n = 8). p- value calculated by two-way matched ANOVA test. i uPAR and int-β1 expression in CTC groups designated as Ki67 Low or Ki67 High ( n = 32). p -value calculated by two-way matched ANOVA. j Ki67 expression in CTC groups designated as uPAR−/int-β1− or uPAR+/int-β1+ ( n = 16). p -value calculated by unpaired t -test. k Circos plot depicting cancer-related gene exon insertions/deletion (in/del) mutations in uPAR+/int-β1+ and uPAR−/int-β1− CTC subsets obtained from BCBM and no BCBM patients. From outer to inner circle : uPAR+/int-β1+ and uPAR−/int β1− CTCs derived from BCBM patient; uPAR+/int β1+ and uPAR−/int-β1− CTCs derived from no BCBM patient. 881/884 in/del mutations found in BCBM subsets, whereas 434/211 in/del in no BCBM subsets. Error bars represent s.e.m

    Article Snippet: Anti-human FITC-CD45 (#304054, dilution 1:200), CD34 (#343504, dilution 1:200), CD105 (#323204, dilution 1:200), CD90 (#328108, dilution 1:200), CD73 (#344016, dilution 1:100), APC-Cy7-CD44 (#103028, dilution 1:100) and BV650-CD44 (#103049, dilution 1:100), BV510-CD24 (#311126, dilution 1:100), APC-int-β1 (#303008, dilution 1:100), AF647-Pan-cytokeratin (#628604, dilution 1:100), PE-PCNA (#307908, dilution 1:100), and AF594-PCNA (#307914, dilution 1:100), BV421-Ki67 (#350506, dilution 1:100), PE-uPAR (#336906, dilution 1:100) from Biolegend.

    Techniques: Derivative Assay, Staining, Isolation, Expressing, Flow Cytometry, Cytometry, Marker

    PARI localization at replication forks and DNA damage sites. (A) HEK 293T (top) and NIH 3T3 (bottom) cells transiently transfected with pCAG-FLAG-PARI and pCAG-PARI-6×His plasmids were immunostained with anti-FLAG and anti-6×His antibodies, respectively. DAPI was used to visualize the nuclei. Scale bars, 5 μm. (B) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI immunostained for FLAG (red) and PCNA (green). Representative images of early, mid-, and late S phase cells are presented. The insets show higher-magnification views of the boxed regions. Scale bars, 5 μm. (C) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI and pulse-labeled with BrdU were stained with anti-FLAG (red) and anti-BrdU (green) antibodies. (D) iPOND analysis of 293T cells transfected with pCAG-FLAG-PARI and pulse-labeled with EdU. Nascent replication forks were captured by the click reaction, and FLAG-PARI and PCNA were detected by Western blotting (WB). The input lanes represent 2.4% of the cell lysates used for the iPOND elution. (E) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI were treated with MMC for 24 h and immunostained for FLAG (green) and PCNA (red). (F) NIH 3T3 cells transiently transfected with plasmids that express FLAG-tagged full-length PARI (top) or PIP box deletion mutant PARIΔPIP (bottom) constructs were stained with anti-FLAG antibody (green). Nuclei were counterstained with DAPI (blue). (G) U2OS cells transfected with a plasmid that expresses GFP-human PARI were laser irradiated (between arrowheads) during S phase (left) or G 1 /G 2 phase (right), and live-cell imaging was carried out. Representative images at the indicated time points are shown. Scale bars, 5 μm.

    Journal: Molecular and Cellular Biology

    Article Title: PARI Regulates Stalled Replication Fork Processing To Maintain Genome Stability upon Replication Stress in Mice

    doi: 10.1128/MCB.00117-17

    Figure Lengend Snippet: PARI localization at replication forks and DNA damage sites. (A) HEK 293T (top) and NIH 3T3 (bottom) cells transiently transfected with pCAG-FLAG-PARI and pCAG-PARI-6×His plasmids were immunostained with anti-FLAG and anti-6×His antibodies, respectively. DAPI was used to visualize the nuclei. Scale bars, 5 μm. (B) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI immunostained for FLAG (red) and PCNA (green). Representative images of early, mid-, and late S phase cells are presented. The insets show higher-magnification views of the boxed regions. Scale bars, 5 μm. (C) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI and pulse-labeled with BrdU were stained with anti-FLAG (red) and anti-BrdU (green) antibodies. (D) iPOND analysis of 293T cells transfected with pCAG-FLAG-PARI and pulse-labeled with EdU. Nascent replication forks were captured by the click reaction, and FLAG-PARI and PCNA were detected by Western blotting (WB). The input lanes represent 2.4% of the cell lysates used for the iPOND elution. (E) NIH 3T3 cells transiently transfected with pCAG-FLAG-PARI were treated with MMC for 24 h and immunostained for FLAG (green) and PCNA (red). (F) NIH 3T3 cells transiently transfected with plasmids that express FLAG-tagged full-length PARI (top) or PIP box deletion mutant PARIΔPIP (bottom) constructs were stained with anti-FLAG antibody (green). Nuclei were counterstained with DAPI (blue). (G) U2OS cells transfected with a plasmid that expresses GFP-human PARI were laser irradiated (between arrowheads) during S phase (left) or G 1 /G 2 phase (right), and live-cell imaging was carried out. Representative images at the indicated time points are shown. Scale bars, 5 μm.

    Article Snippet: SDS-PAGE and Western blotting were carried out according to standard procedures using anti-FLAG polyclonal antibody (1:2,000; Sigma-Aldrich) and anti-PCNA monoclonal antibody (1:1,000; PC10; Biolegend).

    Techniques: Transfection, Labeling, Staining, Western Blot, Mutagenesis, Construct, Plasmid Preparation, Irradiation, Live Cell Imaging