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ATCC hct116

( A ) Example of the immunofluorescence imaging of <t>HCT116</t> cells using antibodies to MLH1. Hoechst staining was used to mark the nucleus. Note the reduced steady-state levels of the G67R MLH1 variant compared to wild-type MLH1. ( B ) The total fluorescent intensity for each of the 69 different MLH1 variants was determined after excluding the non-transfected cells and normalizing the intensities to that for wild-type MLH1. The intensities were then plotted vs. the predicted ΔΔG values. Between 200 and 1,000 cells were included for each quantification. The error bars indicate the standard error of the mean (n = 5 experiments). Each variant is color-coded according to the ClinVar disease category. ( C ) Distribution of steady-state levels by DME category – 0 is loss-of-function in all assays by , 3 represents function in all these assays (for details see the Materials and Methods). Raincloud plot visualization as described in . Colored surface, smoothed density estimate. Gray dots represent means within each DME category, with bars for standard error. ( D ) Distribution of FoldX ΔΔGs across DME categories (as in ( C )). ( E ) FoldX ΔΔGs for all variants tested in this work, indicating their position in the MLH1 sequence. As elsewhere, values above 15 kcal/mol were here set to this value.

Hct116, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 476 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tissue+expression+of+vim/HCT116+VIM+RFP/pmc06837844-70-7-9
Average 99 stars, based on 476 article reviews

hct116 - by Bioz Stars, 2026-10

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1) Product Images from "Computational and cellular studies reveal structural destabilization and degradation of MLH1 variants in Lynch syndrome"

Article Title: Computational and cellular studies reveal structural destabilization and degradation of MLH1 variants in Lynch syndrome

Journal: eLife

doi: 10.7554/eLife.49138

( A ) Example of the immunofluorescence imaging of HCT116 cells using antibodies to MLH1. Hoechst staining was used to mark the nucleus. Note the reduced steady-state levels of the G67R MLH1 variant compared to wild-type MLH1. ( B ) The total fluorescent intensity for each of the 69 different MLH1 variants was determined after excluding the non-transfected cells and normalizing the intensities to that for wild-type MLH1. The intensities were then plotted vs. the predicted ΔΔG values. Between 200 and 1,000 cells were included for each quantification. The error bars indicate the standard error of the mean (n = 5 experiments). Each variant is color-coded according to the ClinVar disease category. ( C ) Distribution of steady-state levels by DME category – 0 is loss-of-function in all assays by , 3 represents function in all these assays (for details see the Materials and Methods). Raincloud plot visualization as described in . Colored surface, smoothed density estimate. Gray dots represent means within each DME category, with bars for standard error. ( D ) Distribution of FoldX ΔΔGs across DME categories (as in ( C )). ( E ) FoldX ΔΔGs for all variants tested in this work, indicating their position in the MLH1 sequence. As elsewhere, values above 15 kcal/mol were here set to this value.

Figure Legend Snippet: ( A ) Example of the immunofluorescence imaging of HCT116 cells using antibodies to MLH1. Hoechst staining was used to mark the nucleus. Note the reduced steady-state levels of the G67R MLH1 variant compared to wild-type MLH1. ( B ) The total fluorescent intensity for each of the 69 different MLH1 variants was determined after excluding the non-transfected cells and normalizing the intensities to that for wild-type MLH1. The intensities were then plotted vs. the predicted ΔΔG values. Between 200 and 1,000 cells were included for each quantification. The error bars indicate the standard error of the mean (n = 5 experiments). Each variant is color-coded according to the ClinVar disease category. ( C ) Distribution of steady-state levels by DME category – 0 is loss-of-function in all assays by , 3 represents function in all these assays (for details see the Materials and Methods). Raincloud plot visualization as described in . Colored surface, smoothed density estimate. Gray dots represent means within each DME category, with bars for standard error. ( D ) Distribution of FoldX ΔΔGs across DME categories (as in ( C )). ( E ) FoldX ΔΔGs for all variants tested in this work, indicating their position in the MLH1 sequence. As elsewhere, values above 15 kcal/mol were here set to this value.

Techniques Used: Immunofluorescence, Imaging, Staining, Variant Assay, Transfection, Sequencing

( A ) HCT116 cells transfected with the indicated variants were analyzed by blotting with antibodies to MLH1. Co-transfection with a plasmid expressing GFP was included to test the transfection efficiencies between the MLH1 variants. β-actin served as a loading control. ( B ) Quantification of blots as in ( A ) normalized to the steady-state level of wild-type (WT) MLH1. The error bars show the standard deviation (n = 3). ( C ) MLH1-transfected HCT116 cells were treated with 25 µg/mL cycloheximide (CHX) for 0, 4, 8 or 12 hr, and lysates were analyzed by blotting using antibodies to MLH1. β-actin was used as a loading control. ( D ) Quantification of blots as in panel ( C ), normalized to the steady-state levels at t = 0 hr. The error bars indicate the standard deviation (n = 3). ( E ) Western blotting with antibodies to MLH1 of whole cell lysates from transfected cells either untreated or treated for 16 hr with 10 µM bortezomib (BZ). Blotting for β-actin was included as a loading control.

Figure Legend Snippet: ( A ) HCT116 cells transfected with the indicated variants were analyzed by blotting with antibodies to MLH1. Co-transfection with a plasmid expressing GFP was included to test the transfection efficiencies between the MLH1 variants. β-actin served as a loading control. ( B ) Quantification of blots as in ( A ) normalized to the steady-state level of wild-type (WT) MLH1. The error bars show the standard deviation (n = 3). ( C ) MLH1-transfected HCT116 cells were treated with 25 µg/mL cycloheximide (CHX) for 0, 4, 8 or 12 hr, and lysates were analyzed by blotting using antibodies to MLH1. β-actin was used as a loading control. ( D ) Quantification of blots as in panel ( C ), normalized to the steady-state levels at t = 0 hr. The error bars indicate the standard deviation (n = 3). ( E ) Western blotting with antibodies to MLH1 of whole cell lysates from transfected cells either untreated or treated for 16 hr with 10 µM bortezomib (BZ). Blotting for β-actin was included as a loading control.

Techniques Used: Transfection, Cotransfection, Plasmid Preparation, Expressing, Control, Standard Deviation, Western Blot

( A ) The levels of endogenous PMS1 and PMS2 were determined by blotting of whole-cell lysates of HCT116 cells transfected with either empty vector or with wild-type MLH1 and treated with 25 µg/mL cycloheximide (CHX) for 0, 4, 8 or 12 hr. The antibodies used were to PMS1 and PMS2, and as a control to MLH1. β-actin served as loading control. ( B ) Quantification of blots as in panel ( A ) normalized to protein levels at 0 hr. The error bars indicate the standard deviation (n = 3). ( C ) The levels of endogenous MLH1, PMS1 and PMS2 were compared by blotting of cell lysates of HCT116 cells either untreated, or treated with cycloheximide (CHX) or with bortezomib (BZ) and CHX. β-actin served as loading control. ( D ) The levels of endogenous PMS1 and PMS2 and transfected MLH1 were compared by western blotting using antibodies to PMS1, PMS2 and MLH1. β-actin served as loading control. ( E ) Quantification of blots as in panel ( C ) normalized to the level of endogenous PMS1 (grey) or PMS2 (red) in untransfected HCT116 cells. The error bars show the standard deviation (n = 3). ( F ) Plotting the levels of the MLH1 variants vs. the levels of endogenous PMS1 (grey) and PMS2 (red). The error bars show the standard deviation (n = 3). ( G ) The levels of MLH1 and YFP-tagged PMS2 were analyzed by SDS-PAGE and blotting of whole-cell lysates of HCT116 cells transfected with the indicated expression plasmids. β-actin was included as loading control. ( H ) Co-transfected PMS2-YFP was immunoprecipitated (IP) using GFP-trap beads, and the precipitated material was analyzed by electrophoresis and blotting. Bortezomib was added to all cultures 16 hr prior to cell lysis to ensure ample amounts of the unstable MLH1 variants.

Figure Legend Snippet: ( A ) The levels of endogenous PMS1 and PMS2 were determined by blotting of whole-cell lysates of HCT116 cells transfected with either empty vector or with wild-type MLH1 and treated with 25 µg/mL cycloheximide (CHX) for 0, 4, 8 or 12 hr. The antibodies used were to PMS1 and PMS2, and as a control to MLH1. β-actin served as loading control. ( B ) Quantification of blots as in panel ( A ) normalized to protein levels at 0 hr. The error bars indicate the standard deviation (n = 3). ( C ) The levels of endogenous MLH1, PMS1 and PMS2 were compared by blotting of cell lysates of HCT116 cells either untreated, or treated with cycloheximide (CHX) or with bortezomib (BZ) and CHX. β-actin served as loading control. ( D ) The levels of endogenous PMS1 and PMS2 and transfected MLH1 were compared by western blotting using antibodies to PMS1, PMS2 and MLH1. β-actin served as loading control. ( E ) Quantification of blots as in panel ( C ) normalized to the level of endogenous PMS1 (grey) or PMS2 (red) in untransfected HCT116 cells. The error bars show the standard deviation (n = 3). ( F ) Plotting the levels of the MLH1 variants vs. the levels of endogenous PMS1 (grey) and PMS2 (red). The error bars show the standard deviation (n = 3). ( G ) The levels of MLH1 and YFP-tagged PMS2 were analyzed by SDS-PAGE and blotting of whole-cell lysates of HCT116 cells transfected with the indicated expression plasmids. β-actin was included as loading control. ( H ) Co-transfected PMS2-YFP was immunoprecipitated (IP) using GFP-trap beads, and the precipitated material was analyzed by electrophoresis and blotting. Bortezomib was added to all cultures 16 hr prior to cell lysis to ensure ample amounts of the unstable MLH1 variants.

Techniques Used: Transfection, Plasmid Preparation, Control, Standard Deviation, Western Blot, SDS Page, Expressing, Immunoprecipitation, Electrophoresis, Lysis

Figure Legend Snippet:

Techniques Used: Recombinant, Software

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