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plko 1 puro constructs  (Addgene inc)


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    Structured Review

    Addgene inc plko 1 puro constructs
    Plko 1 Puro Constructs, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1549 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plasmid+construction/pLKO%2E1+puro+(Plasmid+%238453)/pmc13010945-62-9-16
    Average 96 stars, based on 1549 article reviews
    plko 1 puro constructs - by Bioz Stars, 2026-09
    96/100 stars

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    other:

    Article Title: The evolutionary modifications of a GoLoco motif in the AGS protein facilitate micromere formation in the sea urchin embryo
    Article Snippet: DOI: https://doi.org/10.7554/eLife.100086 23 of 30 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Recombinant DNA reagent Plasmid: SpAGS- GFP Poon et al., 2019; doi:10.1038/s41467-01911560-8 N/A Recombinant DNA reagent Plasmids: SpAGS- dC- term- GFP, SpAGS- dGL1/2/3/4- GFP, SpAGS1111/2222/2134/4234- GFP This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmids: SpAGS- dN- term- GFP This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: SpAGS- mCherry Poon et al., 2019; doi:10.1038/s41467-01911560-8 N/A Recombinant DNA reagent Plasmid: GFP- SpAGS, GFP- EtAGS, GFP- PmAGS, 2x- GFP- SpAGS, 2x- GFPSbAGS This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: GFP- SpAGS4444/GL1GL2/LGNGL/DmGL/EtGL/AGS3GL/PmGL/ S389A/AGS3GL- 3S/A/AGS3GL- GL2GL3 This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: GFP- PmAGS- SpLinker This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: GFP- SpDlg/PmDlg This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: GFP- SpInsc/EtInsc/PmInsc This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: GFP- NuMA This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: mCherry- NuMA This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: GFP- Par3 This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: Vasa- GFP Yajima and Wessel, 2011; doi:10.1242/ dev.054940 N/A Recombinant DNA reagent Plasmid: Vasa- mCherry Uchida and Yajima, 2018; doi:10.1016/j. ydbio.2018.06.015 N/A Recombinant DNA reagent Plasmid: 3xFlag- GFP- SpAGS/SpDlg/SpNuMA, 3xFlag- Vasa- GFP This article N/A See ‘Plasmid construction’ Recombinant DNA reagent Plasmid: 2x- mCherry- EMTB Addgene 26742 Software, algorithm EchinoBase http://www.echinobase. org/Echinobase/ N/A Echinoderm protein sequences Software, algorithm NCBI blast https://blast.ncbi.nlm. nih.gov/Blast.cgi N/A Protein motif search Software, algorithm Clustal Omega https://www.ebi.ac.uk/ Tools/msa/clustalo/ N/A Protein sequence alignment Software, algorithm ImageJ https://imagej.nih. gov/ij/ N/A Quantitative analysis Software, algorithm GraphPad PRISM 8 https://www.graphpad. com/scientific-software/ prism/ N/A Statistical analysis Sequencebased reagent SpAGS- MO NM_001040405.1 Morpholino antisense oligos GGCC CGTT TCAC AAAG CCTT TGTT T Continued on next page Continued Emura, Wavreil et al. eLife 2024;13:RP100086.

    Plasmid Preparation:

    Article Title: Local Chromatin Organization and Net Charge Modulate DNA Glycosylase Efficiency in Live Cells
    Article Snippet: Aptamer Imaging Buffer: 5 mM MgCl2, 0.1 mg/ml Salmon dsDNA (Thermofisher), and 0.1 mg/ml dextran sulphate sodium in 1PBS. .. 10 Plasmid construction: The mEos3.2-C1 plasmid (Addgene #54550), a gift from Michael Davidson and Tao Xu, was used as a backbone to construct hOGG1-mEos3.2 by fusing the hOGG1 sequence (Addgene #18709, provided by David Sidransky) to the N-terminus of mEos3.2 at the AgeI restriction site. .. The piRFP702-N1 plasmid (Addgene #45456), a gift from Vladislav Verkhusha, served as a backbone for creating iRFP702-tagged SRSF2, NPMI, and 15 HP1α constructs by inserting their respective DNA sequences (purchased from Miaolingbio Inc., plasmids #P3758 for SRSF2, #P61047 for NPMI, and #P35426 for HP1α) into the C-terminal region of iRFP702 between SacI and BamHI restriction sites. mEOS3.2-H2B was constructed by inserting their respective DNA sequences (purchased from Miaolingbio Inc., plasmids #P44472 for H2B) into the C-terminal region of mEOS3.2-C1 between SacI and SalI restriction sites.

    Article Title: CPT1A-mediated MFF succinylation promotes stemness maintenance in ovarian cancer stem cells.
    Article Snippet: .. Plasmid construction and lentivirus packaging For shRNA knockdown, shRNA lentivirus vectors were generated by cloning target-specific oligonucleotides into the pLKO.1-TRC shRNA plasmid following themanufacturer’s protocol (Addgene, USA). ..

    Article Title: Supporting Information for WDR20 Prevents Hepatocellular Carcinoma Senescence by Orchestrating the Simultaneous USP12/46-Mediated Deubiquitylation of c-Myc
    Article Snippet: .. Plasmid construction and lentivirus transduction ShRNAs targeting WDR20, WDR48, USP12, USP46 or USP28 were subcloned into pLKO.1-TRC cloning vectors (#10878, Addgene) using the primers listed in SI Appendix, Table S3. .. WDR20 and its corresponding mutants, c-Myc and its corresponding mutants, USP12 and USP46 were subcloned into pCDH-CMV-MCS-EF1-Puro plasmids (#CD510B-1, System Biosciences) with an indicated tag for transient expression using the primers listed in SI Appendix, Table S3.

    Article Title: Utility of the Base Editing System for Introducing Drug-Resistant Gene Mutations Into Human Leukemia Cellular Models
    Article Snippet: Analyses were performed using a CytoVision system (Applied Imaging, Santa Clara, CA). .. Plasmid construction and transfection pCMV_BE4max_P2A_GFP (Addgene #112099) and MLM3636 (Addgene #43860) were provided by David Liu and Keith Joung, respectively. .. Numbers of the potential off-target sites were evaluated by Cas-OFFinder ( http://www.rgenome.net/cas-offinder /) [ ].

    Article Title: The habenula-interpeduncular nucleus-median raphe pathway regulates the outcome of social dominance conflicts in mice.
    Article Snippet: .. Plasmid construction and virus production pEMS2113 (ssAAV-Ple67-emGFP, #49138, Addgene), pAAV-phSyn1(S)-FlpO-bGHpA (#51669, Addgene), pAAV-CAG-fDIO-oGWPRE-SV40pA (#74291, Addgene), pAAV-hSyn-DIO-hM4Di-mCherry (#44362, Addgene), pAAV-hSyn-DIO-hM3Dq-mCherry (#44361, Addgene), and pAAV-hSyn-DIO-mCherry-WPRE-bGHpA (#50459, Addgene) were purchased from Addgene. ..

    Article Title: Selict-seq profiles genome-wide off-target effects in adenosine base editing.
    Article Snippet: HEK293T (ATCC, CRL-11268) and MCF7 (ATCC, HTB-22) cells were separately maintained in Dulbecco’s modified Eagle medium (DMEM) and Minimum Essential Medium (MEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin / streptomycin at 37 ◦C under 5% CO 2 . .. Plasmid construction and sgRNA cloning The ABE8e (TadA-8e V106W) expression plasmid (Addgene, #138495) and ABE7.10 expression plasmid (Addgene, #102919) were purchased and used for transfection. ..

    Construct:

    Article Title: Local Chromatin Organization and Net Charge Modulate DNA Glycosylase Efficiency in Live Cells
    Article Snippet: Aptamer Imaging Buffer: 5 mM MgCl2, 0.1 mg/ml Salmon dsDNA (Thermofisher), and 0.1 mg/ml dextran sulphate sodium in 1PBS. .. 10 Plasmid construction: The mEos3.2-C1 plasmid (Addgene #54550), a gift from Michael Davidson and Tao Xu, was used as a backbone to construct hOGG1-mEos3.2 by fusing the hOGG1 sequence (Addgene #18709, provided by David Sidransky) to the N-terminus of mEos3.2 at the AgeI restriction site. .. The piRFP702-N1 plasmid (Addgene #45456), a gift from Vladislav Verkhusha, served as a backbone for creating iRFP702-tagged SRSF2, NPMI, and 15 HP1α constructs by inserting their respective DNA sequences (purchased from Miaolingbio Inc., plasmids #P3758 for SRSF2, #P61047 for NPMI, and #P35426 for HP1α) into the C-terminal region of iRFP702 between SacI and BamHI restriction sites. mEOS3.2-H2B was constructed by inserting their respective DNA sequences (purchased from Miaolingbio Inc., plasmids #P44472 for H2B) into the C-terminal region of mEOS3.2-C1 between SacI and SalI restriction sites.

    Sequencing:

    Article Title: Local Chromatin Organization and Net Charge Modulate DNA Glycosylase Efficiency in Live Cells
    Article Snippet: Aptamer Imaging Buffer: 5 mM MgCl2, 0.1 mg/ml Salmon dsDNA (Thermofisher), and 0.1 mg/ml dextran sulphate sodium in 1PBS. .. 10 Plasmid construction: The mEos3.2-C1 plasmid (Addgene #54550), a gift from Michael Davidson and Tao Xu, was used as a backbone to construct hOGG1-mEos3.2 by fusing the hOGG1 sequence (Addgene #18709, provided by David Sidransky) to the N-terminus of mEos3.2 at the AgeI restriction site. .. The piRFP702-N1 plasmid (Addgene #45456), a gift from Vladislav Verkhusha, served as a backbone for creating iRFP702-tagged SRSF2, NPMI, and 15 HP1α constructs by inserting their respective DNA sequences (purchased from Miaolingbio Inc., plasmids #P3758 for SRSF2, #P61047 for NPMI, and #P35426 for HP1α) into the C-terminal region of iRFP702 between SacI and BamHI restriction sites. mEOS3.2-H2B was constructed by inserting their respective DNA sequences (purchased from Miaolingbio Inc., plasmids #P44472 for H2B) into the C-terminal region of mEOS3.2-C1 between SacI and SalI restriction sites.

    Article Title: Indole-3-lactic acid suppresses colorectal cancer via metabolic reprogramming.
    Article Snippet: .. Plasmid construction and short hairpin RNA (shRNA) transfection The human HK2 sequence and STAT3 sequence were synthesized by Corues Biotechnology (Nanjing, China) and subsequently subcloned into a pcDNA3.1 vector sourced from Addgene (Cambridge, MA) (Table S8). shRNAs targeting the AHR were procured from Corues Biotechnology, specifically shAHR no.1-TRCN0000021258, shAHR no.2-TRCN0000245285, and no.3-TRCN0000245 286. .. Transfection of shAHR into cells was carried out using Lipofectamine 3000(Thermo Fisher, L3000001), and plasmid transfection was also conducted using the same reagent.

    shRNA:

    Article Title: CPT1A-mediated MFF succinylation promotes stemness maintenance in ovarian cancer stem cells.
    Article Snippet: .. Plasmid construction and lentivirus packaging For shRNA knockdown, shRNA lentivirus vectors were generated by cloning target-specific oligonucleotides into the pLKO.1-TRC shRNA plasmid following themanufacturer’s protocol (Addgene, USA). ..

    Knockdown:

    Article Title: CPT1A-mediated MFF succinylation promotes stemness maintenance in ovarian cancer stem cells.
    Article Snippet: .. Plasmid construction and lentivirus packaging For shRNA knockdown, shRNA lentivirus vectors were generated by cloning target-specific oligonucleotides into the pLKO.1-TRC shRNA plasmid following themanufacturer’s protocol (Addgene, USA). ..

    Generated:

    Article Title: CPT1A-mediated MFF succinylation promotes stemness maintenance in ovarian cancer stem cells.
    Article Snippet: .. Plasmid construction and lentivirus packaging For shRNA knockdown, shRNA lentivirus vectors were generated by cloning target-specific oligonucleotides into the pLKO.1-TRC shRNA plasmid following themanufacturer’s protocol (Addgene, USA). ..

    Cloning:

    Article Title: CPT1A-mediated MFF succinylation promotes stemness maintenance in ovarian cancer stem cells.
    Article Snippet: .. Plasmid construction and lentivirus packaging For shRNA knockdown, shRNA lentivirus vectors were generated by cloning target-specific oligonucleotides into the pLKO.1-TRC shRNA plasmid following themanufacturer’s protocol (Addgene, USA). ..

    Article Title: Supporting Information for WDR20 Prevents Hepatocellular Carcinoma Senescence by Orchestrating the Simultaneous USP12/46-Mediated Deubiquitylation of c-Myc
    Article Snippet: .. Plasmid construction and lentivirus transduction ShRNAs targeting WDR20, WDR48, USP12, USP46 or USP28 were subcloned into pLKO.1-TRC cloning vectors (#10878, Addgene) using the primers listed in SI Appendix, Table S3. .. WDR20 and its corresponding mutants, c-Myc and its corresponding mutants, USP12 and USP46 were subcloned into pCDH-CMV-MCS-EF1-Puro plasmids (#CD510B-1, System Biosciences) with an indicated tag for transient expression using the primers listed in SI Appendix, Table S3.

    Article Title: Selict-seq profiles genome-wide off-target effects in adenosine base editing.
    Article Snippet: HEK293T (ATCC, CRL-11268) and MCF7 (ATCC, HTB-22) cells were separately maintained in Dulbecco’s modified Eagle medium (DMEM) and Minimum Essential Medium (MEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin / streptomycin at 37 ◦C under 5% CO 2 . .. Plasmid construction and sgRNA cloning The ABE8e (TadA-8e V106W) expression plasmid (Addgene, #138495) and ABE7.10 expression plasmid (Addgene, #102919) were purchased and used for transfection. ..

    Transduction:

    Article Title: Supporting Information for WDR20 Prevents Hepatocellular Carcinoma Senescence by Orchestrating the Simultaneous USP12/46-Mediated Deubiquitylation of c-Myc
    Article Snippet: .. Plasmid construction and lentivirus transduction ShRNAs targeting WDR20, WDR48, USP12, USP46 or USP28 were subcloned into pLKO.1-TRC cloning vectors (#10878, Addgene) using the primers listed in SI Appendix, Table S3. .. WDR20 and its corresponding mutants, c-Myc and its corresponding mutants, USP12 and USP46 were subcloned into pCDH-CMV-MCS-EF1-Puro plasmids (#CD510B-1, System Biosciences) with an indicated tag for transient expression using the primers listed in SI Appendix, Table S3.

    Transfection:

    Article Title: Indole-3-lactic acid suppresses colorectal cancer via metabolic reprogramming.
    Article Snippet: .. Plasmid construction and short hairpin RNA (shRNA) transfection The human HK2 sequence and STAT3 sequence were synthesized by Corues Biotechnology (Nanjing, China) and subsequently subcloned into a pcDNA3.1 vector sourced from Addgene (Cambridge, MA) (Table S8). shRNAs targeting the AHR were procured from Corues Biotechnology, specifically shAHR no.1-TRCN0000021258, shAHR no.2-TRCN0000245285, and no.3-TRCN0000245 286. .. Transfection of shAHR into cells was carried out using Lipofectamine 3000(Thermo Fisher, L3000001), and plasmid transfection was also conducted using the same reagent.

    Article Title: Utility of the Base Editing System for Introducing Drug-Resistant Gene Mutations Into Human Leukemia Cellular Models
    Article Snippet: Analyses were performed using a CytoVision system (Applied Imaging, Santa Clara, CA). .. Plasmid construction and transfection pCMV_BE4max_P2A_GFP (Addgene #112099) and MLM3636 (Addgene #43860) were provided by David Liu and Keith Joung, respectively. .. Numbers of the potential off-target sites were evaluated by Cas-OFFinder ( http://www.rgenome.net/cas-offinder /) [ ].

    Article Title: Selict-seq profiles genome-wide off-target effects in adenosine base editing.
    Article Snippet: HEK293T (ATCC, CRL-11268) and MCF7 (ATCC, HTB-22) cells were separately maintained in Dulbecco’s modified Eagle medium (DMEM) and Minimum Essential Medium (MEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin / streptomycin at 37 ◦C under 5% CO 2 . .. Plasmid construction and sgRNA cloning The ABE8e (TadA-8e V106W) expression plasmid (Addgene, #138495) and ABE7.10 expression plasmid (Addgene, #102919) were purchased and used for transfection. ..

    Synthesized:

    Article Title: Indole-3-lactic acid suppresses colorectal cancer via metabolic reprogramming.
    Article Snippet: .. Plasmid construction and short hairpin RNA (shRNA) transfection The human HK2 sequence and STAT3 sequence were synthesized by Corues Biotechnology (Nanjing, China) and subsequently subcloned into a pcDNA3.1 vector sourced from Addgene (Cambridge, MA) (Table S8). shRNAs targeting the AHR were procured from Corues Biotechnology, specifically shAHR no.1-TRCN0000021258, shAHR no.2-TRCN0000245285, and no.3-TRCN0000245 286. .. Transfection of shAHR into cells was carried out using Lipofectamine 3000(Thermo Fisher, L3000001), and plasmid transfection was also conducted using the same reagent.

    Virus:

    Article Title: The habenula-interpeduncular nucleus-median raphe pathway regulates the outcome of social dominance conflicts in mice.
    Article Snippet: .. Plasmid construction and virus production pEMS2113 (ssAAV-Ple67-emGFP, #49138, Addgene), pAAV-phSyn1(S)-FlpO-bGHpA (#51669, Addgene), pAAV-CAG-fDIO-oGWPRE-SV40pA (#74291, Addgene), pAAV-hSyn-DIO-hM4Di-mCherry (#44362, Addgene), pAAV-hSyn-DIO-hM3Dq-mCherry (#44361, Addgene), and pAAV-hSyn-DIO-mCherry-WPRE-bGHpA (#50459, Addgene) were purchased from Addgene. ..

    Expressing:

    Article Title: Selict-seq profiles genome-wide off-target effects in adenosine base editing.
    Article Snippet: HEK293T (ATCC, CRL-11268) and MCF7 (ATCC, HTB-22) cells were separately maintained in Dulbecco’s modified Eagle medium (DMEM) and Minimum Essential Medium (MEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% penicillin / streptomycin at 37 ◦C under 5% CO 2 . .. Plasmid construction and sgRNA cloning The ABE8e (TadA-8e V106W) expression plasmid (Addgene, #138495) and ABE7.10 expression plasmid (Addgene, #102919) were purchased and used for transfection. ..



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    Image Search Results


    KIF23 and myosin heavy chain 9 (MYH9) interact with ubiquitin‐specific protease 7 (USP7). (A) Molecular docking results for KIF23 and MYH9. (B) Co‐immunoprecipitation (Co‐IP) analysis to detect the interaction between KIF23 and MYH9 in CC cells. (C) Immunofluorescence (IF) analysis showing the co‐localisation of KIF23 and MYH9 in CC cells (scale bar: 10 µm). (D) Reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) analysis of MYH9 mRNA levels in CC cells with or without KIF23 KO. (E) Western blotting (WB) analysis of MYH9 protein levels in CC cells with or without KIF23 KO. (F) Nuclear and cytoplasmic fractionation assay to detect MYH9 protein levels in overexpression KIF23 (oe‐ KIF23 ) CC cells. (G) Schematic diagram of the functional domains of KIF23. (H) Co‐IP assay demonstrating the interaction between different KIF23 domains and MYH9 in HEK‐293T cells transfected with the corresponding constructs. (I) Schematic diagram of the functional domains of MYH9. (J) Co‐IP assay demonstrating the interaction between different MYH9 domains and KIF23 in HEK‐293T cells. (K and L) Molecular docking diagrams of MYH9 with USP7 and KIF23 with USP7. (M) Co‐IP analysis in CC cells demonstrating the interactions of MYH9 and KIF23 with USP7. (N and O) Representative confocal images showing the co‐localisation of MYH9 with USP7 and KIF23 with USP7 (scale bar: 10 µm).

    Journal: Clinical and Translational Medicine

    Article Title: Targeting KIF23 inhibits cell proliferation and primary chemoresistance in cervical cancer by inactivating the MYH9/MCM2/PCNA pathway

    doi: 10.1002/ctm2.70652

    Figure Lengend Snippet: KIF23 and myosin heavy chain 9 (MYH9) interact with ubiquitin‐specific protease 7 (USP7). (A) Molecular docking results for KIF23 and MYH9. (B) Co‐immunoprecipitation (Co‐IP) analysis to detect the interaction between KIF23 and MYH9 in CC cells. (C) Immunofluorescence (IF) analysis showing the co‐localisation of KIF23 and MYH9 in CC cells (scale bar: 10 µm). (D) Reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) analysis of MYH9 mRNA levels in CC cells with or without KIF23 KO. (E) Western blotting (WB) analysis of MYH9 protein levels in CC cells with or without KIF23 KO. (F) Nuclear and cytoplasmic fractionation assay to detect MYH9 protein levels in overexpression KIF23 (oe‐ KIF23 ) CC cells. (G) Schematic diagram of the functional domains of KIF23. (H) Co‐IP assay demonstrating the interaction between different KIF23 domains and MYH9 in HEK‐293T cells transfected with the corresponding constructs. (I) Schematic diagram of the functional domains of MYH9. (J) Co‐IP assay demonstrating the interaction between different MYH9 domains and KIF23 in HEK‐293T cells. (K and L) Molecular docking diagrams of MYH9 with USP7 and KIF23 with USP7. (M) Co‐IP analysis in CC cells demonstrating the interactions of MYH9 and KIF23 with USP7. (N and O) Representative confocal images showing the co‐localisation of MYH9 with USP7 and KIF23 with USP7 (scale bar: 10 µm).

    Article Snippet: CC cells were transfected with plasmid constructs encoding USP7 (Miaoling Biology), MYH9 (Vigene Biosciences), MCM2 (Miaoling Biology) and USP15 (Miaoling Biology) using Lipofectamine 3000 (model L2000‐015; Invitrogen) as per the manufacturer's protocols.

    Techniques: Ubiquitin Proteomics, Immunoprecipitation, Co-Immunoprecipitation Assay, Immunofluorescence, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Fractionation, Over Expression, Functional Assay, Transfection, Construct

    KIF23 induces K48‐linked deubiquitination of MYH9 through recruitment of USP7. (A) Western blotting (WB) analysis of MYH9 protein stability at different time points following cycloheximide (CHX) treatment in control and KIF23 KO groups. (B) CHX chase assay examining the regulation of MYH9 protein stability by USP7 overexpression (oe‐ USP7 ). (C) Effect of MG132 treatment (12 h) on MYH9 protein stability in KIF23 KO CC cells and their controls. (D) Co‐IP and WB analyses of the effect of KIF23 KO on MYH9 ubiquitination levels after 12 h of MG132 treatment. (E) Co‐IP and WB assays assessing the effects of control, KIF23 KO and KIF23 KO combined with USP7 overexpression (KO‐ KIF23 + oe‐ USP7 ) on MYH9 ubiquitination after 12 h of MG132 treatment in SIHA (left) and C33A (right) cells.

    Journal: Clinical and Translational Medicine

    Article Title: Targeting KIF23 inhibits cell proliferation and primary chemoresistance in cervical cancer by inactivating the MYH9/MCM2/PCNA pathway

    doi: 10.1002/ctm2.70652

    Figure Lengend Snippet: KIF23 induces K48‐linked deubiquitination of MYH9 through recruitment of USP7. (A) Western blotting (WB) analysis of MYH9 protein stability at different time points following cycloheximide (CHX) treatment in control and KIF23 KO groups. (B) CHX chase assay examining the regulation of MYH9 protein stability by USP7 overexpression (oe‐ USP7 ). (C) Effect of MG132 treatment (12 h) on MYH9 protein stability in KIF23 KO CC cells and their controls. (D) Co‐IP and WB analyses of the effect of KIF23 KO on MYH9 ubiquitination levels after 12 h of MG132 treatment. (E) Co‐IP and WB assays assessing the effects of control, KIF23 KO and KIF23 KO combined with USP7 overexpression (KO‐ KIF23 + oe‐ USP7 ) on MYH9 ubiquitination after 12 h of MG132 treatment in SIHA (left) and C33A (right) cells.

    Article Snippet: CC cells were transfected with plasmid constructs encoding USP7 (Miaoling Biology), MYH9 (Vigene Biosciences), MCM2 (Miaoling Biology) and USP15 (Miaoling Biology) using Lipofectamine 3000 (model L2000‐015; Invitrogen) as per the manufacturer's protocols.

    Techniques: Western Blot, Control, Over Expression, Co-Immunoprecipitation Assay, Ubiquitin Proteomics