human embryonic kidney 293 hek293 cell lines  (ATCC)


Bioz Verified Symbol ATCC is a verified supplier
Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC human embryonic kidney 293 hek293 cell lines
    ( A ) <t>HEK293</t> cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.
    Human Embryonic Kidney 293 Hek293 Cell Lines, supplied by ATCC, 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/human embryonic kidney 293 hek293 cell lines/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney 293 hek293 cell lines - by Bioz Stars, 2023-09
    86/100 stars

    Images

    1) Product Images from "The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery"

    Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

    Journal: eLife

    doi: 10.7554/eLife.86976

    ( A ) HEK293 cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.
    Figure Legend Snippet: ( A ) HEK293 cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.

    Techniques Used: Western Blot, Incubation

    ( A–C ) HeLa cells expressing CFP-MASTL were treated without or with ionizing radiation (IR). Cells were analyzed by direct fluorescence for CFP-MASTL expression, and by immunofluorescence for the phosphorylation of ATM/ATR substrates. Quantifications are shown in panels B and C (>10 cells/time point). ( D, E ) HEK293 cells were treated without ( D ) or with ( E ) 10 mM hydroxyurea (HU) for 2 hr. These cells were then treated with cycloheximide (CHX, 20 μg/ml) at time 0 to block protein synthesis, and analyzed by immunoblotting for the protein stability of MASTL and α-tubulin.
    Figure Legend Snippet: ( A–C ) HeLa cells expressing CFP-MASTL were treated without or with ionizing radiation (IR). Cells were analyzed by direct fluorescence for CFP-MASTL expression, and by immunofluorescence for the phosphorylation of ATM/ATR substrates. Quantifications are shown in panels B and C (>10 cells/time point). ( D, E ) HEK293 cells were treated without ( D ) or with ( E ) 10 mM hydroxyurea (HU) for 2 hr. These cells were then treated with cycloheximide (CHX, 20 μg/ml) at time 0 to block protein synthesis, and analyzed by immunoblotting for the protein stability of MASTL and α-tubulin.

    Techniques Used: Expressing, Fluorescence, Immunofluorescence, Blocking Assay, Western Blot

    ( A ) The sequence alignment of the conserved E6AP Ser-218 motif in human, mouse, and Xenopus . ( B ) A phospho-specific antibody recognizing E6AP Ser-218 was generated, as described in Materials and methods. WT or E6AP knockout (KO) HEK293 cells were treated without or with 10 mM hydroxyurea (HU) for 4 hr, and analyzed by immunoblotting for phospho-E6AP Ser-218, E6AP, and α-tubulin. ( C ) HeLa cells were treated without or with 0.5 μM doxorubicin (DOX) and 5 μM KU55933 (ATMi), as indicated, for 1 hr, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-E6AP Ser-218, and α-tubulin. ( D ) HeLa cells were transfected with HA-tagged WT, S218A, or S218D E6AP. Cell lysates were harvested for immunoprecipitation (IP) assays. The input, MASTL IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL and HA. ( E ) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX for 3 hr, and harvested for IP assays. The input, HA IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL, phospho-E6AP Ser-218, and HA. ( F ) E6AP KO HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX, incubated as indicated, and harvested for immunoblotting for MASTL and α-tubulin.
    Figure Legend Snippet: ( A ) The sequence alignment of the conserved E6AP Ser-218 motif in human, mouse, and Xenopus . ( B ) A phospho-specific antibody recognizing E6AP Ser-218 was generated, as described in Materials and methods. WT or E6AP knockout (KO) HEK293 cells were treated without or with 10 mM hydroxyurea (HU) for 4 hr, and analyzed by immunoblotting for phospho-E6AP Ser-218, E6AP, and α-tubulin. ( C ) HeLa cells were treated without or with 0.5 μM doxorubicin (DOX) and 5 μM KU55933 (ATMi), as indicated, for 1 hr, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-E6AP Ser-218, and α-tubulin. ( D ) HeLa cells were transfected with HA-tagged WT, S218A, or S218D E6AP. Cell lysates were harvested for immunoprecipitation (IP) assays. The input, MASTL IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL and HA. ( E ) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX for 3 hr, and harvested for IP assays. The input, HA IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL, phospho-E6AP Ser-218, and HA. ( F ) E6AP KO HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX, incubated as indicated, and harvested for immunoblotting for MASTL and α-tubulin.

    Techniques Used: Sequencing, Generated, Knock-Out, Western Blot, Transfection, Immunoprecipitation, Incubation

    ( A ) WT or E6AP knockout HEK293 cells were treated without or with doxorubicin (DOX, 0.5 μM) for 4 hr. Cells were harvested and analyzed by immunoblotting for phospho-E6AP Ser-218 and α-tubulin. ( B ) HEK293 cells were treated without or with DOX (0.5 μM), or ATM inhibitor (KU55933, 10 μM), for 4 hr, and analyzed by immunoblotting. ( C ) HeLa cells were treated with hydroxyurea (HU, 10 mM) combined with ATM/ATR inhibitor (caffeine, 4 mM) or ATM inhibitor (KU55933, 10 μM), for 12 hr, as indicated. Cells were harvested and analyzed by immunoblotting.
    Figure Legend Snippet: ( A ) WT or E6AP knockout HEK293 cells were treated without or with doxorubicin (DOX, 0.5 μM) for 4 hr. Cells were harvested and analyzed by immunoblotting for phospho-E6AP Ser-218 and α-tubulin. ( B ) HEK293 cells were treated without or with DOX (0.5 μM), or ATM inhibitor (KU55933, 10 μM), for 4 hr, and analyzed by immunoblotting. ( C ) HeLa cells were treated with hydroxyurea (HU, 10 mM) combined with ATM/ATR inhibitor (caffeine, 4 mM) or ATM inhibitor (KU55933, 10 μM), for 12 hr, as indicated. Cells were harvested and analyzed by immunoblotting.

    Techniques Used: Knock-Out, Western Blot

    ( A ) E6AP knockout (KO) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in . Cells were treated with 0.1 μM etoposide (ETO) for 18 hr, and released in fresh medium for recovery. Cells were then harvested at the indicated time points (after the removal of ETO) for immunofluorescence (IF) using an anti-phospho-Aurora A/B/C antibody. The activation of Aurora phosphorylation (shown in red) and chromosome condensation (in blue) indicated mitosis. The percentages of cells in mitosis were quantified manually and shown. The mean values and standard deviations were calculated from three experiments. An unpaired two-tailed Student’s t test was used to determine the statistical significance (**p<0.01, n>500 cell numbers/measurement). ( B ) E6AP KO HeLa cells expressing HA-tagged WT or S218A E6AP, as in panel A, were treated with 2 mM hydroxyurea (HU) for 18 hr. Cells were then released in fresh medium, and incubated as indicated, for recovery. Cell cycle progression was analyzed by fluorescence-activated cell sorting (FACS). ( C ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells. Cells were treated without or with 0.1 μM ETO for 18 hr, released in fresh medium for recovery, and incubated as indicated. Cells were analyzed by immunoblotting for phospho-cyclin-dependent kinase (CDK) substrates and histone H3. ( D ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells, as in panel C. Cells were treated without or with 1 μM CPT for 90 min, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-SMC1 Ser-957, and α-tubulin.
    Figure Legend Snippet: ( A ) E6AP knockout (KO) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in . Cells were treated with 0.1 μM etoposide (ETO) for 18 hr, and released in fresh medium for recovery. Cells were then harvested at the indicated time points (after the removal of ETO) for immunofluorescence (IF) using an anti-phospho-Aurora A/B/C antibody. The activation of Aurora phosphorylation (shown in red) and chromosome condensation (in blue) indicated mitosis. The percentages of cells in mitosis were quantified manually and shown. The mean values and standard deviations were calculated from three experiments. An unpaired two-tailed Student’s t test was used to determine the statistical significance (**p<0.01, n>500 cell numbers/measurement). ( B ) E6AP KO HeLa cells expressing HA-tagged WT or S218A E6AP, as in panel A, were treated with 2 mM hydroxyurea (HU) for 18 hr. Cells were then released in fresh medium, and incubated as indicated, for recovery. Cell cycle progression was analyzed by fluorescence-activated cell sorting (FACS). ( C ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells. Cells were treated without or with 0.1 μM ETO for 18 hr, released in fresh medium for recovery, and incubated as indicated. Cells were analyzed by immunoblotting for phospho-cyclin-dependent kinase (CDK) substrates and histone H3. ( D ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells, as in panel C. Cells were treated without or with 1 μM CPT for 90 min, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-SMC1 Ser-957, and α-tubulin.

    Techniques Used: Knock-Out, Transfection, Immunofluorescence, Activation Assay, Two Tailed Test, Expressing, Incubation, Fluorescence, FACS, Western Blot

    human embryonic kidney epithelial cell line 293  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC human embryonic kidney epithelial cell line 293
    Human Embryonic Kidney Epithelial Cell Line 293, supplied by ATCC, 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/human embryonic kidney epithelial cell line 293/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney epithelial cell line 293 - by Bioz Stars, 2023-09
    86/100 stars

    Images

    human embryonic kidney hek 293 cell line  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC human embryonic kidney hek 293 cell line
    Human Embryonic Kidney Hek 293 Cell Line, supplied by ATCC, 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/human embryonic kidney hek 293 cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney hek 293 cell line - by Bioz Stars, 2023-09
    86/100 stars

    Images

    embryonic kidney cell line 293 t  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC embryonic kidney cell line 293 t
    Embryonic Kidney Cell Line 293 T, supplied by ATCC, 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/embryonic kidney cell line 293 t/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    embryonic kidney cell line 293 t - by Bioz Stars, 2023-09
    86/100 stars

    Images

    embryonic kidney 293 hek293 cell line  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC embryonic kidney 293 hek293 cell line
    Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in <t>HEK293</t> and <t>HEK</t> SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.
    Embryonic Kidney 293 Hek293 Cell Line, supplied by ATCC, 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/embryonic kidney 293 hek293 cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    embryonic kidney 293 hek293 cell line - by Bioz Stars, 2023-09
    86/100 stars

    Images

    1) Product Images from "In skeletal muscle and neural crest cells, SMCHD1 regulates biological pathways relevant for Bosma syndrome and facioscapulohumeral dystrophy phenotype"

    Article Title: In skeletal muscle and neural crest cells, SMCHD1 regulates biological pathways relevant for Bosma syndrome and facioscapulohumeral dystrophy phenotype

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkad523

    Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in HEK293 and HEK SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.
    Figure Legend Snippet: Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in HEK293 and HEK SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.

    Techniques Used: Construct, Methylation, Binding Assay, Clone Assay, Luciferase, Expressing, Transfection, Plasmid Preparation, Activity Assay, MANN-WHITNEY, Mutagenesis, Sequencing, Flow Cytometry

    embryonic kidney cell line 293 t  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC embryonic kidney cell line 293 t
    Embryonic Kidney Cell Line 293 T, supplied by ATCC, 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/embryonic kidney cell line 293 t/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    embryonic kidney cell line 293 t - by Bioz Stars, 2023-09
    86/100 stars

    Images

    human embryonic kidney hek 293 cell line  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC human embryonic kidney hek 293 cell line
    Human Embryonic Kidney Hek 293 Cell Line, supplied by ATCC, 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/human embryonic kidney hek 293 cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney hek 293 cell line - by Bioz Stars, 2023-09
    86/100 stars

    Images

    embryonic kidney cell line hek 293  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC embryonic kidney cell line hek 293
    Embryonic Kidney Cell Line Hek 293, supplied by ATCC, 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/embryonic kidney cell line hek 293/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    embryonic kidney cell line hek 293 - by Bioz Stars, 2023-09
    86/100 stars

    Images

    293 t embryonic kidney cell line  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC 293 t embryonic kidney cell line
    293 T Embryonic Kidney Cell Line, supplied by ATCC, 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/293 t embryonic kidney cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    293 t embryonic kidney cell line - by Bioz Stars, 2023-09
    86/100 stars

    Images

    human embryonic kidney hek 293 crl 1573 tm cell line  (ATCC)


    Bioz Verified Symbol ATCC is a verified supplier
    Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    ATCC human embryonic kidney hek 293 crl 1573 tm cell line
    Human Embryonic Kidney Hek 293 Crl 1573 Tm Cell Line, supplied by ATCC, 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/human embryonic kidney hek 293 crl 1573 tm cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney hek 293 crl 1573 tm cell line - by Bioz Stars, 2023-09
    86/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    ATCC human embryonic kidney 293 hek293 cell lines
    ( A ) <t>HEK293</t> cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.
    Human Embryonic Kidney 293 Hek293 Cell Lines, supplied by ATCC, 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/human embryonic kidney 293 hek293 cell lines/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney 293 hek293 cell lines - by Bioz Stars, 2023-09
    86/100 stars
      Buy from Supplier

    86
    ATCC human embryonic kidney epithelial cell line 293
    ( A ) <t>HEK293</t> cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.
    Human Embryonic Kidney Epithelial Cell Line 293, supplied by ATCC, 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/human embryonic kidney epithelial cell line 293/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney epithelial cell line 293 - by Bioz Stars, 2023-09
    86/100 stars
      Buy from Supplier

    86
    ATCC human embryonic kidney hek 293 cell line
    ( A ) <t>HEK293</t> cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.
    Human Embryonic Kidney Hek 293 Cell Line, supplied by ATCC, 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/human embryonic kidney hek 293 cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney hek 293 cell line - by Bioz Stars, 2023-09
    86/100 stars
      Buy from Supplier

    86
    ATCC embryonic kidney cell line 293 t
    ( A ) <t>HEK293</t> cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.
    Embryonic Kidney Cell Line 293 T, supplied by ATCC, 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/embryonic kidney cell line 293 t/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    embryonic kidney cell line 293 t - by Bioz Stars, 2023-09
    86/100 stars
      Buy from Supplier

    86
    ATCC embryonic kidney 293 hek293 cell line
    Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in <t>HEK293</t> and <t>HEK</t> SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.
    Embryonic Kidney 293 Hek293 Cell Line, supplied by ATCC, 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/embryonic kidney 293 hek293 cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    embryonic kidney 293 hek293 cell line - by Bioz Stars, 2023-09
    86/100 stars
      Buy from Supplier

    86
    ATCC embryonic kidney cell line hek 293
    Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in <t>HEK293</t> and <t>HEK</t> SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.
    Embryonic Kidney Cell Line Hek 293, supplied by ATCC, 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/embryonic kidney cell line hek 293/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    embryonic kidney cell line hek 293 - by Bioz Stars, 2023-09
    86/100 stars
      Buy from Supplier

    86
    ATCC 293 t embryonic kidney cell line
    Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in <t>HEK293</t> and <t>HEK</t> SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.
    293 T Embryonic Kidney Cell Line, supplied by ATCC, 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/293 t embryonic kidney cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    293 t embryonic kidney cell line - by Bioz Stars, 2023-09
    86/100 stars
      Buy from Supplier

    86
    ATCC human embryonic kidney hek 293 crl 1573 tm cell line
    Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in <t>HEK293</t> and <t>HEK</t> SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.
    Human Embryonic Kidney Hek 293 Crl 1573 Tm Cell Line, supplied by ATCC, 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/human embryonic kidney hek 293 crl 1573 tm cell line/product/ATCC
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human embryonic kidney hek 293 crl 1573 tm cell line - by Bioz Stars, 2023-09
    86/100 stars
      Buy from Supplier

    Image Search Results


    ( A ) HEK293 cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.

    Journal: eLife

    Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

    doi: 10.7554/eLife.86976

    Figure Lengend Snippet: ( A ) HEK293 cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.

    Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

    Techniques: Western Blot, Incubation

    ( A–C ) HeLa cells expressing CFP-MASTL were treated without or with ionizing radiation (IR). Cells were analyzed by direct fluorescence for CFP-MASTL expression, and by immunofluorescence for the phosphorylation of ATM/ATR substrates. Quantifications are shown in panels B and C (>10 cells/time point). ( D, E ) HEK293 cells were treated without ( D ) or with ( E ) 10 mM hydroxyurea (HU) for 2 hr. These cells were then treated with cycloheximide (CHX, 20 μg/ml) at time 0 to block protein synthesis, and analyzed by immunoblotting for the protein stability of MASTL and α-tubulin.

    Journal: eLife

    Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

    doi: 10.7554/eLife.86976

    Figure Lengend Snippet: ( A–C ) HeLa cells expressing CFP-MASTL were treated without or with ionizing radiation (IR). Cells were analyzed by direct fluorescence for CFP-MASTL expression, and by immunofluorescence for the phosphorylation of ATM/ATR substrates. Quantifications are shown in panels B and C (>10 cells/time point). ( D, E ) HEK293 cells were treated without ( D ) or with ( E ) 10 mM hydroxyurea (HU) for 2 hr. These cells were then treated with cycloheximide (CHX, 20 μg/ml) at time 0 to block protein synthesis, and analyzed by immunoblotting for the protein stability of MASTL and α-tubulin.

    Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

    Techniques: Expressing, Fluorescence, Immunofluorescence, Blocking Assay, Western Blot

    ( A ) The sequence alignment of the conserved E6AP Ser-218 motif in human, mouse, and Xenopus . ( B ) A phospho-specific antibody recognizing E6AP Ser-218 was generated, as described in Materials and methods. WT or E6AP knockout (KO) HEK293 cells were treated without or with 10 mM hydroxyurea (HU) for 4 hr, and analyzed by immunoblotting for phospho-E6AP Ser-218, E6AP, and α-tubulin. ( C ) HeLa cells were treated without or with 0.5 μM doxorubicin (DOX) and 5 μM KU55933 (ATMi), as indicated, for 1 hr, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-E6AP Ser-218, and α-tubulin. ( D ) HeLa cells were transfected with HA-tagged WT, S218A, or S218D E6AP. Cell lysates were harvested for immunoprecipitation (IP) assays. The input, MASTL IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL and HA. ( E ) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX for 3 hr, and harvested for IP assays. The input, HA IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL, phospho-E6AP Ser-218, and HA. ( F ) E6AP KO HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX, incubated as indicated, and harvested for immunoblotting for MASTL and α-tubulin.

    Journal: eLife

    Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

    doi: 10.7554/eLife.86976

    Figure Lengend Snippet: ( A ) The sequence alignment of the conserved E6AP Ser-218 motif in human, mouse, and Xenopus . ( B ) A phospho-specific antibody recognizing E6AP Ser-218 was generated, as described in Materials and methods. WT or E6AP knockout (KO) HEK293 cells were treated without or with 10 mM hydroxyurea (HU) for 4 hr, and analyzed by immunoblotting for phospho-E6AP Ser-218, E6AP, and α-tubulin. ( C ) HeLa cells were treated without or with 0.5 μM doxorubicin (DOX) and 5 μM KU55933 (ATMi), as indicated, for 1 hr, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-E6AP Ser-218, and α-tubulin. ( D ) HeLa cells were transfected with HA-tagged WT, S218A, or S218D E6AP. Cell lysates were harvested for immunoprecipitation (IP) assays. The input, MASTL IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL and HA. ( E ) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX for 3 hr, and harvested for IP assays. The input, HA IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL, phospho-E6AP Ser-218, and HA. ( F ) E6AP KO HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX, incubated as indicated, and harvested for immunoblotting for MASTL and α-tubulin.

    Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

    Techniques: Sequencing, Generated, Knock-Out, Western Blot, Transfection, Immunoprecipitation, Incubation

    ( A ) WT or E6AP knockout HEK293 cells were treated without or with doxorubicin (DOX, 0.5 μM) for 4 hr. Cells were harvested and analyzed by immunoblotting for phospho-E6AP Ser-218 and α-tubulin. ( B ) HEK293 cells were treated without or with DOX (0.5 μM), or ATM inhibitor (KU55933, 10 μM), for 4 hr, and analyzed by immunoblotting. ( C ) HeLa cells were treated with hydroxyurea (HU, 10 mM) combined with ATM/ATR inhibitor (caffeine, 4 mM) or ATM inhibitor (KU55933, 10 μM), for 12 hr, as indicated. Cells were harvested and analyzed by immunoblotting.

    Journal: eLife

    Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

    doi: 10.7554/eLife.86976

    Figure Lengend Snippet: ( A ) WT or E6AP knockout HEK293 cells were treated without or with doxorubicin (DOX, 0.5 μM) for 4 hr. Cells were harvested and analyzed by immunoblotting for phospho-E6AP Ser-218 and α-tubulin. ( B ) HEK293 cells were treated without or with DOX (0.5 μM), or ATM inhibitor (KU55933, 10 μM), for 4 hr, and analyzed by immunoblotting. ( C ) HeLa cells were treated with hydroxyurea (HU, 10 mM) combined with ATM/ATR inhibitor (caffeine, 4 mM) or ATM inhibitor (KU55933, 10 μM), for 12 hr, as indicated. Cells were harvested and analyzed by immunoblotting.

    Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

    Techniques: Knock-Out, Western Blot

    ( A ) E6AP knockout (KO) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in . Cells were treated with 0.1 μM etoposide (ETO) for 18 hr, and released in fresh medium for recovery. Cells were then harvested at the indicated time points (after the removal of ETO) for immunofluorescence (IF) using an anti-phospho-Aurora A/B/C antibody. The activation of Aurora phosphorylation (shown in red) and chromosome condensation (in blue) indicated mitosis. The percentages of cells in mitosis were quantified manually and shown. The mean values and standard deviations were calculated from three experiments. An unpaired two-tailed Student’s t test was used to determine the statistical significance (**p<0.01, n>500 cell numbers/measurement). ( B ) E6AP KO HeLa cells expressing HA-tagged WT or S218A E6AP, as in panel A, were treated with 2 mM hydroxyurea (HU) for 18 hr. Cells were then released in fresh medium, and incubated as indicated, for recovery. Cell cycle progression was analyzed by fluorescence-activated cell sorting (FACS). ( C ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells. Cells were treated without or with 0.1 μM ETO for 18 hr, released in fresh medium for recovery, and incubated as indicated. Cells were analyzed by immunoblotting for phospho-cyclin-dependent kinase (CDK) substrates and histone H3. ( D ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells, as in panel C. Cells were treated without or with 1 μM CPT for 90 min, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-SMC1 Ser-957, and α-tubulin.

    Journal: eLife

    Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

    doi: 10.7554/eLife.86976

    Figure Lengend Snippet: ( A ) E6AP knockout (KO) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in . Cells were treated with 0.1 μM etoposide (ETO) for 18 hr, and released in fresh medium for recovery. Cells were then harvested at the indicated time points (after the removal of ETO) for immunofluorescence (IF) using an anti-phospho-Aurora A/B/C antibody. The activation of Aurora phosphorylation (shown in red) and chromosome condensation (in blue) indicated mitosis. The percentages of cells in mitosis were quantified manually and shown. The mean values and standard deviations were calculated from three experiments. An unpaired two-tailed Student’s t test was used to determine the statistical significance (**p<0.01, n>500 cell numbers/measurement). ( B ) E6AP KO HeLa cells expressing HA-tagged WT or S218A E6AP, as in panel A, were treated with 2 mM hydroxyurea (HU) for 18 hr. Cells were then released in fresh medium, and incubated as indicated, for recovery. Cell cycle progression was analyzed by fluorescence-activated cell sorting (FACS). ( C ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells. Cells were treated without or with 0.1 μM ETO for 18 hr, released in fresh medium for recovery, and incubated as indicated. Cells were analyzed by immunoblotting for phospho-cyclin-dependent kinase (CDK) substrates and histone H3. ( D ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells, as in panel C. Cells were treated without or with 1 μM CPT for 90 min, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-SMC1 Ser-957, and α-tubulin.

    Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

    Techniques: Knock-Out, Transfection, Immunofluorescence, Activation Assay, Two Tailed Test, Expressing, Incubation, Fluorescence, FACS, Western Blot

    Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in HEK293 and HEK SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.

    Journal: Nucleic Acids Research

    Article Title: In skeletal muscle and neural crest cells, SMCHD1 regulates biological pathways relevant for Bosma syndrome and facioscapulohumeral dystrophy phenotype

    doi: 10.1093/nar/gkad523

    Figure Lengend Snippet: Depending on the genomic context, SMCHD1 contributes to gene silencing or protects against position effects. For the different pGL3 constructs, fragments corresponding to the regions that are differentially methylated ( A – C ) or corresponding to SMCHD1 binding sites ( D ) were cloned downstream of the luciferase reporter gene in vectors lacking an enhancer (pGL3 promoter). Firefly luciferase expression was determined 48 h post-transfection of the different constructs in HEK293 and HEK SMCHD1 KO cells. The pGL3 control vector was used as a transfection control. Firely luciferase levels were normalized to expression of the Renilla luciferase used as transfection control. Values corresponding to the normalized luciferase activity (expressed in relative luminescence units, RLUs) are the average of three independent assays, each realized as technical triplicates ( n = 9). Error bars represent standard error. Statistical significance was determined using a Mann–Whitney test (**** P -value <0.00001, *** P -value <0.0001, ** P -value <0.001, * P -value = 0.01). (A) For the D4Z4 macrosatellite, regions that are differentially methylated in patients carrying a mutation in SMCHD1 were tested (DR1, 5P; a scheme of the D4Z4 repeat is presented in ). (B) The DR1 sequence contains 31 CG sites. Fragments corresponding to CG1–10, CG10–20 and CG21–31 or deleted of 10 of these CGs (ΔCG1–10, ΔCG10–20 and ΔCG21–31) were tested. (C) H OX gene DMRs. (D) Putative SMCHD1 binding sites overlapping or not with CTCF binding sites. ( E , F ) For evaluation of protection against position effect, we used a vector carrying a hygromycin resistance gene and an eGFP reporter gene. Sequences to be tested are cloned downstream of the eGFP gene and transfected into HEK or HEK KO cells. Stable eGFP expression was measured by flow cytometry (FACS) for an extended period of time in cells grown in the presence of hygromycin B. Representative spectra of the % of eGFP-positive cells are presented. For each condition, eGFP expression was compared to values obtained in cells transfected with the empty vector (pCMV). In HEK SMCHD1 KO cells, eGFP expression was also measured 72 h after transfection of an SMCHD1 expression vector (gray curves). (E) D4Z4 (left upper panel), DR1–5P (right upper panel), DR1 (left lower panel) and 5P (right lower panel). (F) Results obtained for the HOXA13 (left) and HOXC4/5/6 (right) DMRs.

    Article Snippet: The human embryonic kidney 293 (HEK293) cell line (CRL-1573) was obtained from ATCC.

    Techniques: Construct, Methylation, Binding Assay, Clone Assay, Luciferase, Expressing, Transfection, Plasmid Preparation, Activity Assay, MANN-WHITNEY, Mutagenesis, Sequencing, Flow Cytometry