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Fluxion Biosciences human embryonic kidney hek293 cells
Patch-clamp analysis of cells treated with miR-502-3p. (A) Sweep plots showing the gamma-aminobutyric acid (GABA) current in the human-gamma-aminobutyric acid receptor A-α1/β2/γ2L (hGABAA-α1/β2/γ2L) <t>human</t> <t>embryonic</t> <t>kidney</t> <t>(HEK)</t> recombinant cells transfected with miR-502-3p agomiRs and miR-502-3p antagomiRs. The endogenous GABA channel was recorded at 300 μm GABA concentration for 4 seconds using the IonFlux system. (B) Quantification of GABA current in the parental, scramble control, miR-502-3p agomiRs, and antagomiR treated cells. The GABA current was reduced in agomiR transfected cells, while it was significantly increased in the miR-502-3p antagomiR transfected cells compared with scramble control. Experiments were performed a minimum 8 times in each group. The P values < 0.05 were considered statistically significant. miR: MicroRNA.
Human Embryonic Kidney Hek293 Cells, supplied by Fluxion Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "MicroRNA-502-3p regulates GABAergic synapse function in hippocampal neurons"

Article Title: MicroRNA-502-3p regulates GABAergic synapse function in hippocampal neurons

Journal: Neural Regeneration Research

doi: 10.4103/NRR.NRR-D-23-01064

Patch-clamp analysis of cells treated with miR-502-3p. (A) Sweep plots showing the gamma-aminobutyric acid (GABA) current in the human-gamma-aminobutyric acid receptor A-α1/β2/γ2L (hGABAA-α1/β2/γ2L) human embryonic kidney (HEK) recombinant cells transfected with miR-502-3p agomiRs and miR-502-3p antagomiRs. The endogenous GABA channel was recorded at 300 μm GABA concentration for 4 seconds using the IonFlux system. (B) Quantification of GABA current in the parental, scramble control, miR-502-3p agomiRs, and antagomiR treated cells. The GABA current was reduced in agomiR transfected cells, while it was significantly increased in the miR-502-3p antagomiR transfected cells compared with scramble control. Experiments were performed a minimum 8 times in each group. The P values < 0.05 were considered statistically significant. miR: MicroRNA.
Figure Legend Snippet: Patch-clamp analysis of cells treated with miR-502-3p. (A) Sweep plots showing the gamma-aminobutyric acid (GABA) current in the human-gamma-aminobutyric acid receptor A-α1/β2/γ2L (hGABAA-α1/β2/γ2L) human embryonic kidney (HEK) recombinant cells transfected with miR-502-3p agomiRs and miR-502-3p antagomiRs. The endogenous GABA channel was recorded at 300 μm GABA concentration for 4 seconds using the IonFlux system. (B) Quantification of GABA current in the parental, scramble control, miR-502-3p agomiRs, and antagomiR treated cells. The GABA current was reduced in agomiR transfected cells, while it was significantly increased in the miR-502-3p antagomiR transfected cells compared with scramble control. Experiments were performed a minimum 8 times in each group. The P values < 0.05 were considered statistically significant. miR: MicroRNA.

Techniques Used: Patch Clamp, Recombinant, Transfection, Concentration Assay

hek 293 a human embryonic kidney cell line  (ATCC)


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    ATCC hek 293 a human embryonic kidney cell line
    Phosphorylation-independent MAb production and epitope mapping. (A) Reactivity of MAbs with TDP-43-Flag recombinant protein of about 46 kDa (lane 1) overexpressed in <t>HEK</t> <t>293</t> cells by Western blot. Anti-Flag MAb (Flag) was used as the positive control antibody. Leucine-rich repeat-containing protein 15 (LRRC15)-His fusion protein (lane 2) served as the negative control, which was revealed by anti-His MAb (HIS). (B) Schematic representation of the TDP-43 fragments employed for epitope mapping. (C) Mapping of TDP-43 domains with epitopes recognized by the 10 generated MAbs. This mapping was performed across the entire length of TDP-43 using immunoblot and indirect peptide ELISA. RRM, RNA-recognition motif; NLS, bipartite nuclear localization signal; NES, nuclear export signal.
    Hek 293 A Human 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
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    1) Product Images from "Production and characterization of novel monoclonal antibodies against pathological human TDP-43 proteins"

    Article Title: Production and characterization of novel monoclonal antibodies against pathological human TDP-43 proteins

    Journal: Journal of Neuropathology and Experimental Neurology

    doi: 10.1093/jnen/nlae042

    Phosphorylation-independent MAb production and epitope mapping. (A) Reactivity of MAbs with TDP-43-Flag recombinant protein of about 46 kDa (lane 1) overexpressed in HEK 293 cells by Western blot. Anti-Flag MAb (Flag) was used as the positive control antibody. Leucine-rich repeat-containing protein 15 (LRRC15)-His fusion protein (lane 2) served as the negative control, which was revealed by anti-His MAb (HIS). (B) Schematic representation of the TDP-43 fragments employed for epitope mapping. (C) Mapping of TDP-43 domains with epitopes recognized by the 10 generated MAbs. This mapping was performed across the entire length of TDP-43 using immunoblot and indirect peptide ELISA. RRM, RNA-recognition motif; NLS, bipartite nuclear localization signal; NES, nuclear export signal.
    Figure Legend Snippet: Phosphorylation-independent MAb production and epitope mapping. (A) Reactivity of MAbs with TDP-43-Flag recombinant protein of about 46 kDa (lane 1) overexpressed in HEK 293 cells by Western blot. Anti-Flag MAb (Flag) was used as the positive control antibody. Leucine-rich repeat-containing protein 15 (LRRC15)-His fusion protein (lane 2) served as the negative control, which was revealed by anti-His MAb (HIS). (B) Schematic representation of the TDP-43 fragments employed for epitope mapping. (C) Mapping of TDP-43 domains with epitopes recognized by the 10 generated MAbs. This mapping was performed across the entire length of TDP-43 using immunoblot and indirect peptide ELISA. RRM, RNA-recognition motif; NLS, bipartite nuclear localization signal; NES, nuclear export signal.

    Techniques Used: Recombinant, Western Blot, Positive Control, Negative Control, Generated, Peptide ELISA

    hek human embryonic kidney 293 f cells  (Thermo Fisher)


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    Thermo Fisher hek human embryonic kidney 293 f cells
    Hek Human Embryonic Kidney 293 F Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    embryonic kidney 293 hek293 cells  (ATCC)


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    ATCC embryonic kidney 293 hek293 cells
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    human embryonic kidney hek293 cells  (ATCC)


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    ATCC human embryonic kidney hek293 cells
    ABCB1 cell surface expression. Flow cytometry histograms of <t>HEK293</t> cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).
    Human Embryonic Kidney Hek293 Cells, 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
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    1) Product Images from "Effect of ABCB1 most frequent polymorphisms on the accumulation of bictegravir in recombinant HEK293 cell lines"

    Article Title: Effect of ABCB1 most frequent polymorphisms on the accumulation of bictegravir in recombinant HEK293 cell lines

    Journal: Scientific Reports

    doi: 10.1038/s41598-024-66809-0

    ABCB1 cell surface expression. Flow cytometry histograms of HEK293 cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).
    Figure Legend Snippet: ABCB1 cell surface expression. Flow cytometry histograms of HEK293 cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).

    Techniques Used: Expressing, Flow Cytometry, Transfection, Plasmid Preparation, Incubation, Control

    Model functionality assays using the ABCB1 substrate Rhodamine123 with or without the ABCB1 inhibitor Zosuquidar. Protein normalized intracellular fluorescence measured after incubation of ( a ) HEK control and HEK 1199A and ( b ) HEK control , HEK WT , HEK CGT and HEK TTT with the fluorescent ABCB1 substrate Rhodamine123 (Rh123), in the absence (left) and in the presence (right) of Zosuquidar (Zos). Intensity of fluorescence is reported in arbitrary units. Results from a single experiment (N = 1 for ( a ) and N = 1 for ( b )) reported as mean ± standard error (number of technical replicates performed per condition (n) = 3 for ( a ); n = 5 for ( b )). Statistical analysis reported: ANOVA-2, after natural logarithmic transformation for ( b ). **p < 0.01; ****p < 0.0001.
    Figure Legend Snippet: Model functionality assays using the ABCB1 substrate Rhodamine123 with or without the ABCB1 inhibitor Zosuquidar. Protein normalized intracellular fluorescence measured after incubation of ( a ) HEK control and HEK 1199A and ( b ) HEK control , HEK WT , HEK CGT and HEK TTT with the fluorescent ABCB1 substrate Rhodamine123 (Rh123), in the absence (left) and in the presence (right) of Zosuquidar (Zos). Intensity of fluorescence is reported in arbitrary units. Results from a single experiment (N = 1 for ( a ) and N = 1 for ( b )) reported as mean ± standard error (number of technical replicates performed per condition (n) = 3 for ( a ); n = 5 for ( b )). Statistical analysis reported: ANOVA-2, after natural logarithmic transformation for ( b ). **p < 0.01; ****p < 0.0001.

    Techniques Used: Fluorescence, Incubation, Control, Transformation Assay

    Intracellular protein-normalized bictegravir (BIC) concentrations in HEK control , HEK WT and HEK 1199A cells after 2 h of incubation at several dose levels. Results of three experiments were pooled (N = 3). Protein normalized BIC intracellular concentrations are reported as fold-change of the mean of the protein normalized BIC intracellular concentration observed at the lowest BIC exposition concentration for HEK control (number of technical replicates performed per condition and per experiment = 3) . Results reported as mean ± standard error. Statistical analysis reported: ANOVA-2 performed on pooled Ln-transformed data. **p < 0.01 ***p < 0.001 ****p < 0.0001.
    Figure Legend Snippet: Intracellular protein-normalized bictegravir (BIC) concentrations in HEK control , HEK WT and HEK 1199A cells after 2 h of incubation at several dose levels. Results of three experiments were pooled (N = 3). Protein normalized BIC intracellular concentrations are reported as fold-change of the mean of the protein normalized BIC intracellular concentration observed at the lowest BIC exposition concentration for HEK control (number of technical replicates performed per condition and per experiment = 3) . Results reported as mean ± standard error. Statistical analysis reported: ANOVA-2 performed on pooled Ln-transformed data. **p < 0.01 ***p < 0.001 ****p < 0.0001.

    Techniques Used: Control, Incubation, Concentration Assay, Transformation Assay

    Intracellular protein-normalized bictegravir (BIC) concentrations in HEK control , HEK WT , HEK CGT and HEK TTT cells after 2 h of incubation at several dose levels. Results of three experiments were pooled (N = 3). Protein normalized BIC intracellular concentrations are reported as fold-change of the mean of the protein normalized BIC intracellular concentration observed at the lowest BIC exposition concentration for HEK control (number of technical replicates performed per condition and per experiment = 3) . Results reported as mean ± standard error. Statistical analysis reported: ANOVA-2 performed on pooled Ln-transformed data. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
    Figure Legend Snippet: Intracellular protein-normalized bictegravir (BIC) concentrations in HEK control , HEK WT , HEK CGT and HEK TTT cells after 2 h of incubation at several dose levels. Results of three experiments were pooled (N = 3). Protein normalized BIC intracellular concentrations are reported as fold-change of the mean of the protein normalized BIC intracellular concentration observed at the lowest BIC exposition concentration for HEK control (number of technical replicates performed per condition and per experiment = 3) . Results reported as mean ± standard error. Statistical analysis reported: ANOVA-2 performed on pooled Ln-transformed data. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Techniques Used: Control, Incubation, Concentration Assay, Transformation Assay

    human embryonic kidney hek 293 cells  (Thermo Fisher)


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    Thermo Fisher human embryonic kidney hek 293 cells
    Human Embryonic Kidney Hek 293 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    embryonic kidney hek293 cells  (Thermo Fisher)


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    Thermo Fisher embryonic kidney hek293 cells
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    human embryonic kidney hek293 cells  (ATCC)


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    ATCC human embryonic kidney hek293 cells
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    embryonic kidney hek 293 t cells  (Thermo Fisher)


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    Thermo Fisher embryonic kidney hek 293 t cells
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    human embryonic kidney hek 293 t cells  (ATCC)


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    ATCC human embryonic kidney hek 293 t cells
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    Fluxion Biosciences human embryonic kidney hek293 cells
    Patch-clamp analysis of cells treated with miR-502-3p. (A) Sweep plots showing the gamma-aminobutyric acid (GABA) current in the human-gamma-aminobutyric acid receptor A-α1/β2/γ2L (hGABAA-α1/β2/γ2L) <t>human</t> <t>embryonic</t> <t>kidney</t> <t>(HEK)</t> recombinant cells transfected with miR-502-3p agomiRs and miR-502-3p antagomiRs. The endogenous GABA channel was recorded at 300 μm GABA concentration for 4 seconds using the IonFlux system. (B) Quantification of GABA current in the parental, scramble control, miR-502-3p agomiRs, and antagomiR treated cells. The GABA current was reduced in agomiR transfected cells, while it was significantly increased in the miR-502-3p antagomiR transfected cells compared with scramble control. Experiments were performed a minimum 8 times in each group. The P values < 0.05 were considered statistically significant. miR: MicroRNA.
    Human Embryonic Kidney Hek293 Cells, supplied by Fluxion Biosciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC hek 293 a human embryonic kidney cell line
    Phosphorylation-independent MAb production and epitope mapping. (A) Reactivity of MAbs with TDP-43-Flag recombinant protein of about 46 kDa (lane 1) overexpressed in <t>HEK</t> <t>293</t> cells by Western blot. Anti-Flag MAb (Flag) was used as the positive control antibody. Leucine-rich repeat-containing protein 15 (LRRC15)-His fusion protein (lane 2) served as the negative control, which was revealed by anti-His MAb (HIS). (B) Schematic representation of the TDP-43 fragments employed for epitope mapping. (C) Mapping of TDP-43 domains with epitopes recognized by the 10 generated MAbs. This mapping was performed across the entire length of TDP-43 using immunoblot and indirect peptide ELISA. RRM, RNA-recognition motif; NLS, bipartite nuclear localization signal; NES, nuclear export signal.
    Hek 293 A Human 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
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    Thermo Fisher hek human embryonic kidney 293 f cells
    Phosphorylation-independent MAb production and epitope mapping. (A) Reactivity of MAbs with TDP-43-Flag recombinant protein of about 46 kDa (lane 1) overexpressed in <t>HEK</t> <t>293</t> cells by Western blot. Anti-Flag MAb (Flag) was used as the positive control antibody. Leucine-rich repeat-containing protein 15 (LRRC15)-His fusion protein (lane 2) served as the negative control, which was revealed by anti-His MAb (HIS). (B) Schematic representation of the TDP-43 fragments employed for epitope mapping. (C) Mapping of TDP-43 domains with epitopes recognized by the 10 generated MAbs. This mapping was performed across the entire length of TDP-43 using immunoblot and indirect peptide ELISA. RRM, RNA-recognition motif; NLS, bipartite nuclear localization signal; NES, nuclear export signal.
    Hek Human Embryonic Kidney 293 F Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC embryonic kidney 293 hek293 cells
    Phosphorylation-independent MAb production and epitope mapping. (A) Reactivity of MAbs with TDP-43-Flag recombinant protein of about 46 kDa (lane 1) overexpressed in <t>HEK</t> <t>293</t> cells by Western blot. Anti-Flag MAb (Flag) was used as the positive control antibody. Leucine-rich repeat-containing protein 15 (LRRC15)-His fusion protein (lane 2) served as the negative control, which was revealed by anti-His MAb (HIS). (B) Schematic representation of the TDP-43 fragments employed for epitope mapping. (C) Mapping of TDP-43 domains with epitopes recognized by the 10 generated MAbs. This mapping was performed across the entire length of TDP-43 using immunoblot and indirect peptide ELISA. RRM, RNA-recognition motif; NLS, bipartite nuclear localization signal; NES, nuclear export signal.
    Embryonic Kidney 293 Hek293 Cells, 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
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    ATCC human embryonic kidney hek293 cells
    ABCB1 cell surface expression. Flow cytometry histograms of <t>HEK293</t> cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).
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    Thermo Fisher human embryonic kidney hek 293 cells
    ABCB1 cell surface expression. Flow cytometry histograms of <t>HEK293</t> cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).
    Human Embryonic Kidney Hek 293 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher embryonic kidney hek293 cells
    ABCB1 cell surface expression. Flow cytometry histograms of <t>HEK293</t> cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).
    Embryonic Kidney Hek293 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher embryonic kidney hek 293 t cells
    ABCB1 cell surface expression. Flow cytometry histograms of <t>HEK293</t> cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).
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    ATCC human embryonic kidney hek 293 t cells
    ABCB1 cell surface expression. Flow cytometry histograms of <t>HEK293</t> cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).
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    Patch-clamp analysis of cells treated with miR-502-3p. (A) Sweep plots showing the gamma-aminobutyric acid (GABA) current in the human-gamma-aminobutyric acid receptor A-α1/β2/γ2L (hGABAA-α1/β2/γ2L) human embryonic kidney (HEK) recombinant cells transfected with miR-502-3p agomiRs and miR-502-3p antagomiRs. The endogenous GABA channel was recorded at 300 μm GABA concentration for 4 seconds using the IonFlux system. (B) Quantification of GABA current in the parental, scramble control, miR-502-3p agomiRs, and antagomiR treated cells. The GABA current was reduced in agomiR transfected cells, while it was significantly increased in the miR-502-3p antagomiR transfected cells compared with scramble control. Experiments were performed a minimum 8 times in each group. The P values < 0.05 were considered statistically significant. miR: MicroRNA.

    Journal: Neural Regeneration Research

    Article Title: MicroRNA-502-3p regulates GABAergic synapse function in hippocampal neurons

    doi: 10.4103/NRR.NRR-D-23-01064

    Figure Lengend Snippet: Patch-clamp analysis of cells treated with miR-502-3p. (A) Sweep plots showing the gamma-aminobutyric acid (GABA) current in the human-gamma-aminobutyric acid receptor A-α1/β2/γ2L (hGABAA-α1/β2/γ2L) human embryonic kidney (HEK) recombinant cells transfected with miR-502-3p agomiRs and miR-502-3p antagomiRs. The endogenous GABA channel was recorded at 300 μm GABA concentration for 4 seconds using the IonFlux system. (B) Quantification of GABA current in the parental, scramble control, miR-502-3p agomiRs, and antagomiR treated cells. The GABA current was reduced in agomiR transfected cells, while it was significantly increased in the miR-502-3p antagomiR transfected cells compared with scramble control. Experiments were performed a minimum 8 times in each group. The P values < 0.05 were considered statistically significant. miR: MicroRNA.

    Article Snippet: Briefly, human embryonic kidney (HEK293) cells expressing human-gamma-aminobutyric acid receptors hGABAA-HEK293α1/β2/γ2L (Cat# 909-0069, Fluxion Biosciences Inc.) were cultured in 175 cm 2 filter-top flasks containing DMEM/ F12 glutamax, G-418, hygromycin B, and puromycin at 37°C and 5% CO 2 .

    Techniques: Patch Clamp, Recombinant, Transfection, Concentration Assay

    Phosphorylation-independent MAb production and epitope mapping. (A) Reactivity of MAbs with TDP-43-Flag recombinant protein of about 46 kDa (lane 1) overexpressed in HEK 293 cells by Western blot. Anti-Flag MAb (Flag) was used as the positive control antibody. Leucine-rich repeat-containing protein 15 (LRRC15)-His fusion protein (lane 2) served as the negative control, which was revealed by anti-His MAb (HIS). (B) Schematic representation of the TDP-43 fragments employed for epitope mapping. (C) Mapping of TDP-43 domains with epitopes recognized by the 10 generated MAbs. This mapping was performed across the entire length of TDP-43 using immunoblot and indirect peptide ELISA. RRM, RNA-recognition motif; NLS, bipartite nuclear localization signal; NES, nuclear export signal.

    Journal: Journal of Neuropathology and Experimental Neurology

    Article Title: Production and characterization of novel monoclonal antibodies against pathological human TDP-43 proteins

    doi: 10.1093/jnen/nlae042

    Figure Lengend Snippet: Phosphorylation-independent MAb production and epitope mapping. (A) Reactivity of MAbs with TDP-43-Flag recombinant protein of about 46 kDa (lane 1) overexpressed in HEK 293 cells by Western blot. Anti-Flag MAb (Flag) was used as the positive control antibody. Leucine-rich repeat-containing protein 15 (LRRC15)-His fusion protein (lane 2) served as the negative control, which was revealed by anti-His MAb (HIS). (B) Schematic representation of the TDP-43 fragments employed for epitope mapping. (C) Mapping of TDP-43 domains with epitopes recognized by the 10 generated MAbs. This mapping was performed across the entire length of TDP-43 using immunoblot and indirect peptide ELISA. RRM, RNA-recognition motif; NLS, bipartite nuclear localization signal; NES, nuclear export signal.

    Article Snippet: Two cell lines, SH-SY5Y (a human neuroblastoma cell line) and HEK 293 (a human embryonic kidney cell line), were sourced from the American Type Culture Collection (ATCC, Manassas, VA).

    Techniques: Recombinant, Western Blot, Positive Control, Negative Control, Generated, Peptide ELISA

    ABCB1 cell surface expression. Flow cytometry histograms of HEK293 cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).

    Journal: Scientific Reports

    Article Title: Effect of ABCB1 most frequent polymorphisms on the accumulation of bictegravir in recombinant HEK293 cell lines

    doi: 10.1038/s41598-024-66809-0

    Figure Lengend Snippet: ABCB1 cell surface expression. Flow cytometry histograms of HEK293 cells transfected with ( a ) the empty pcDNA3.1 vector, ABCB1 WT or ABCB1 1199A and ( b ) the empty pcDNA3.1 vector, ABCB1 WT , ABCB1 CGT or ABCB1 TTT . Cells were incubated in the absence of antibody (autofluorescence, orange), with isotype control (blue) or with FITC anti-ABCB1 antibody (red).

    Article Snippet: Briefly, human embryonic kidney (HEK293) cells were purchased from American Type Culture Collection and transfected with plasmids carrying either wild-type ABCB1 1199G-1236C-2677G-3435C (thereafter named HEK WT ) or several ABCB1 variant combinations obtained using site-directed mutagenesis, 1199A-1236C-2677G-3435C (thereafter named HEK 1199A ), 1199G-1236C-2677G-3435T (thereafter named HEK CGT ) and 1199G-1236T-2677T-3435T (thereafter named HEK TTT ); or an empty vector (thereafter named HEK control ).

    Techniques: Expressing, Flow Cytometry, Transfection, Plasmid Preparation, Incubation, Control

    Model functionality assays using the ABCB1 substrate Rhodamine123 with or without the ABCB1 inhibitor Zosuquidar. Protein normalized intracellular fluorescence measured after incubation of ( a ) HEK control and HEK 1199A and ( b ) HEK control , HEK WT , HEK CGT and HEK TTT with the fluorescent ABCB1 substrate Rhodamine123 (Rh123), in the absence (left) and in the presence (right) of Zosuquidar (Zos). Intensity of fluorescence is reported in arbitrary units. Results from a single experiment (N = 1 for ( a ) and N = 1 for ( b )) reported as mean ± standard error (number of technical replicates performed per condition (n) = 3 for ( a ); n = 5 for ( b )). Statistical analysis reported: ANOVA-2, after natural logarithmic transformation for ( b ). **p < 0.01; ****p < 0.0001.

    Journal: Scientific Reports

    Article Title: Effect of ABCB1 most frequent polymorphisms on the accumulation of bictegravir in recombinant HEK293 cell lines

    doi: 10.1038/s41598-024-66809-0

    Figure Lengend Snippet: Model functionality assays using the ABCB1 substrate Rhodamine123 with or without the ABCB1 inhibitor Zosuquidar. Protein normalized intracellular fluorescence measured after incubation of ( a ) HEK control and HEK 1199A and ( b ) HEK control , HEK WT , HEK CGT and HEK TTT with the fluorescent ABCB1 substrate Rhodamine123 (Rh123), in the absence (left) and in the presence (right) of Zosuquidar (Zos). Intensity of fluorescence is reported in arbitrary units. Results from a single experiment (N = 1 for ( a ) and N = 1 for ( b )) reported as mean ± standard error (number of technical replicates performed per condition (n) = 3 for ( a ); n = 5 for ( b )). Statistical analysis reported: ANOVA-2, after natural logarithmic transformation for ( b ). **p < 0.01; ****p < 0.0001.

    Article Snippet: Briefly, human embryonic kidney (HEK293) cells were purchased from American Type Culture Collection and transfected with plasmids carrying either wild-type ABCB1 1199G-1236C-2677G-3435C (thereafter named HEK WT ) or several ABCB1 variant combinations obtained using site-directed mutagenesis, 1199A-1236C-2677G-3435C (thereafter named HEK 1199A ), 1199G-1236C-2677G-3435T (thereafter named HEK CGT ) and 1199G-1236T-2677T-3435T (thereafter named HEK TTT ); or an empty vector (thereafter named HEK control ).

    Techniques: Fluorescence, Incubation, Control, Transformation Assay

    Intracellular protein-normalized bictegravir (BIC) concentrations in HEK control , HEK WT and HEK 1199A cells after 2 h of incubation at several dose levels. Results of three experiments were pooled (N = 3). Protein normalized BIC intracellular concentrations are reported as fold-change of the mean of the protein normalized BIC intracellular concentration observed at the lowest BIC exposition concentration for HEK control (number of technical replicates performed per condition and per experiment = 3) . Results reported as mean ± standard error. Statistical analysis reported: ANOVA-2 performed on pooled Ln-transformed data. **p < 0.01 ***p < 0.001 ****p < 0.0001.

    Journal: Scientific Reports

    Article Title: Effect of ABCB1 most frequent polymorphisms on the accumulation of bictegravir in recombinant HEK293 cell lines

    doi: 10.1038/s41598-024-66809-0

    Figure Lengend Snippet: Intracellular protein-normalized bictegravir (BIC) concentrations in HEK control , HEK WT and HEK 1199A cells after 2 h of incubation at several dose levels. Results of three experiments were pooled (N = 3). Protein normalized BIC intracellular concentrations are reported as fold-change of the mean of the protein normalized BIC intracellular concentration observed at the lowest BIC exposition concentration for HEK control (number of technical replicates performed per condition and per experiment = 3) . Results reported as mean ± standard error. Statistical analysis reported: ANOVA-2 performed on pooled Ln-transformed data. **p < 0.01 ***p < 0.001 ****p < 0.0001.

    Article Snippet: Briefly, human embryonic kidney (HEK293) cells were purchased from American Type Culture Collection and transfected with plasmids carrying either wild-type ABCB1 1199G-1236C-2677G-3435C (thereafter named HEK WT ) or several ABCB1 variant combinations obtained using site-directed mutagenesis, 1199A-1236C-2677G-3435C (thereafter named HEK 1199A ), 1199G-1236C-2677G-3435T (thereafter named HEK CGT ) and 1199G-1236T-2677T-3435T (thereafter named HEK TTT ); or an empty vector (thereafter named HEK control ).

    Techniques: Control, Incubation, Concentration Assay, Transformation Assay

    Intracellular protein-normalized bictegravir (BIC) concentrations in HEK control , HEK WT , HEK CGT and HEK TTT cells after 2 h of incubation at several dose levels. Results of three experiments were pooled (N = 3). Protein normalized BIC intracellular concentrations are reported as fold-change of the mean of the protein normalized BIC intracellular concentration observed at the lowest BIC exposition concentration for HEK control (number of technical replicates performed per condition and per experiment = 3) . Results reported as mean ± standard error. Statistical analysis reported: ANOVA-2 performed on pooled Ln-transformed data. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Journal: Scientific Reports

    Article Title: Effect of ABCB1 most frequent polymorphisms on the accumulation of bictegravir in recombinant HEK293 cell lines

    doi: 10.1038/s41598-024-66809-0

    Figure Lengend Snippet: Intracellular protein-normalized bictegravir (BIC) concentrations in HEK control , HEK WT , HEK CGT and HEK TTT cells after 2 h of incubation at several dose levels. Results of three experiments were pooled (N = 3). Protein normalized BIC intracellular concentrations are reported as fold-change of the mean of the protein normalized BIC intracellular concentration observed at the lowest BIC exposition concentration for HEK control (number of technical replicates performed per condition and per experiment = 3) . Results reported as mean ± standard error. Statistical analysis reported: ANOVA-2 performed on pooled Ln-transformed data. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

    Article Snippet: Briefly, human embryonic kidney (HEK293) cells were purchased from American Type Culture Collection and transfected with plasmids carrying either wild-type ABCB1 1199G-1236C-2677G-3435C (thereafter named HEK WT ) or several ABCB1 variant combinations obtained using site-directed mutagenesis, 1199A-1236C-2677G-3435C (thereafter named HEK 1199A ), 1199G-1236C-2677G-3435T (thereafter named HEK CGT ) and 1199G-1236T-2677T-3435T (thereafter named HEK TTT ); or an empty vector (thereafter named HEK control ).

    Techniques: Control, Incubation, Concentration Assay, Transformation Assay