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  • 94
    Cell Signaling Technology Inc rabbit polyclonal anti tdp43
    (A) Structure of <t>TDP43</t> RRM1 and RRM2, generated in FirstGlance in Jmol (PDB:4BS2), deposited by . TDP43 (blue) is shown in complex with UG-repeat RNA (green), highlighting the R151-D247 salt bridge (red). (B) TDP43 schematic depicting the RRM1-RRM2 salt bridge and F147-F149. (C) EMSA of increasing concentrations (12 fmol–4 pmol) of recombinant TDP43 variants mixed with labeled (UG) 12 (100 pM). (D, F, and H) Increasing concentrations (0.01–25 μM) of TDP43(WT) (D), TDP43(R151A) (F), and TDP43(D247A) (H) were added to labeled (UG) 12 (100 pM). (E, G, and I) K d and h were calculated from 3 independent replicates for TDP43(WT) (E), TDP43(R151A) (G), and TDP43(D247A) (I). (J and K) EMSA of TDP43 variants at increasing protein concentrations (0.5–16 pmol) incubated with 100 pM of labeled (GC) 12 (J) or (AT) 12 (K). (L and M) RNA IP of EGFP-tagged TDP43(WT), TDP43(F147L, F149L), TDP43(R151A), or TDP43(D247A) expressed in HEK293T cells. The abundance of bound TARDBP (L) or MALAT1 (M) transcripts was measured by qRT-PCR. ****p < 0.0001, one-way ANOVA with Dunnett’s test. Arrowheads in (C), (D), (F), (H), (J), and (K) indicate protein-DNA complexes, while arrows point to unbound oligonucleotides. For (E), (G), and (I) K d and h were determined by nonlinear least-squares regression; #p < 0.0001 for comparison with TDP43(WT) by extra sum-of-squares F test. For (L) and (M), data were pooled from 3 independent replicates. All plots show mean ± SEM.
    Rabbit Polyclonal Anti Tdp43, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti tdp43/product/Cell Signaling Technology Inc
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    86
    Thermo Fisher a 260
    (A) Structure of <t>TDP43</t> RRM1 and RRM2, generated in FirstGlance in Jmol (PDB:4BS2), deposited by . TDP43 (blue) is shown in complex with UG-repeat RNA (green), highlighting the R151-D247 salt bridge (red). (B) TDP43 schematic depicting the RRM1-RRM2 salt bridge and F147-F149. (C) EMSA of increasing concentrations (12 fmol–4 pmol) of recombinant TDP43 variants mixed with labeled (UG) 12 (100 pM). (D, F, and H) Increasing concentrations (0.01–25 μM) of TDP43(WT) (D), TDP43(R151A) (F), and TDP43(D247A) (H) were added to labeled (UG) 12 (100 pM). (E, G, and I) K d and h were calculated from 3 independent replicates for TDP43(WT) (E), TDP43(R151A) (G), and TDP43(D247A) (I). (J and K) EMSA of TDP43 variants at increasing protein concentrations (0.5–16 pmol) incubated with 100 pM of labeled (GC) 12 (J) or (AT) 12 (K). (L and M) RNA IP of EGFP-tagged TDP43(WT), TDP43(F147L, F149L), TDP43(R151A), or TDP43(D247A) expressed in HEK293T cells. The abundance of bound TARDBP (L) or MALAT1 (M) transcripts was measured by qRT-PCR. ****p < 0.0001, one-way ANOVA with Dunnett’s test. Arrowheads in (C), (D), (F), (H), (J), and (K) indicate protein-DNA complexes, while arrows point to unbound oligonucleotides. For (E), (G), and (I) K d and h were determined by nonlinear least-squares regression; #p < 0.0001 for comparison with TDP43(WT) by extra sum-of-squares F test. For (L) and (M), data were pooled from 3 independent replicates. All plots show mean ± SEM.
    A 260, 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
    https://www.bioz.com/result/a 260/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
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    a 260 - by Bioz Stars, 2023-01
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    86
    Varian Medical a 260
    (A) Structure of <t>TDP43</t> RRM1 and RRM2, generated in FirstGlance in Jmol (PDB:4BS2), deposited by . TDP43 (blue) is shown in complex with UG-repeat RNA (green), highlighting the R151-D247 salt bridge (red). (B) TDP43 schematic depicting the RRM1-RRM2 salt bridge and F147-F149. (C) EMSA of increasing concentrations (12 fmol–4 pmol) of recombinant TDP43 variants mixed with labeled (UG) 12 (100 pM). (D, F, and H) Increasing concentrations (0.01–25 μM) of TDP43(WT) (D), TDP43(R151A) (F), and TDP43(D247A) (H) were added to labeled (UG) 12 (100 pM). (E, G, and I) K d and h were calculated from 3 independent replicates for TDP43(WT) (E), TDP43(R151A) (G), and TDP43(D247A) (I). (J and K) EMSA of TDP43 variants at increasing protein concentrations (0.5–16 pmol) incubated with 100 pM of labeled (GC) 12 (J) or (AT) 12 (K). (L and M) RNA IP of EGFP-tagged TDP43(WT), TDP43(F147L, F149L), TDP43(R151A), or TDP43(D247A) expressed in HEK293T cells. The abundance of bound TARDBP (L) or MALAT1 (M) transcripts was measured by qRT-PCR. ****p < 0.0001, one-way ANOVA with Dunnett’s test. Arrowheads in (C), (D), (F), (H), (J), and (K) indicate protein-DNA complexes, while arrows point to unbound oligonucleotides. For (E), (G), and (I) K d and h were determined by nonlinear least-squares regression; #p < 0.0001 for comparison with TDP43(WT) by extra sum-of-squares F test. For (L) and (M), data were pooled from 3 independent replicates. All plots show mean ± SEM.
    A 260, supplied by Varian Medical, 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/a 260/product/Varian Medical
    Average 86 stars, based on 1 article reviews
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    a 260 - by Bioz Stars, 2023-01
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    86
    Agilent technologies a 260
    (A) Structure of <t>TDP43</t> RRM1 and RRM2, generated in FirstGlance in Jmol (PDB:4BS2), deposited by . TDP43 (blue) is shown in complex with UG-repeat RNA (green), highlighting the R151-D247 salt bridge (red). (B) TDP43 schematic depicting the RRM1-RRM2 salt bridge and F147-F149. (C) EMSA of increasing concentrations (12 fmol–4 pmol) of recombinant TDP43 variants mixed with labeled (UG) 12 (100 pM). (D, F, and H) Increasing concentrations (0.01–25 μM) of TDP43(WT) (D), TDP43(R151A) (F), and TDP43(D247A) (H) were added to labeled (UG) 12 (100 pM). (E, G, and I) K d and h were calculated from 3 independent replicates for TDP43(WT) (E), TDP43(R151A) (G), and TDP43(D247A) (I). (J and K) EMSA of TDP43 variants at increasing protein concentrations (0.5–16 pmol) incubated with 100 pM of labeled (GC) 12 (J) or (AT) 12 (K). (L and M) RNA IP of EGFP-tagged TDP43(WT), TDP43(F147L, F149L), TDP43(R151A), or TDP43(D247A) expressed in HEK293T cells. The abundance of bound TARDBP (L) or MALAT1 (M) transcripts was measured by qRT-PCR. ****p < 0.0001, one-way ANOVA with Dunnett’s test. Arrowheads in (C), (D), (F), (H), (J), and (K) indicate protein-DNA complexes, while arrows point to unbound oligonucleotides. For (E), (G), and (I) K d and h were determined by nonlinear least-squares regression; #p < 0.0001 for comparison with TDP43(WT) by extra sum-of-squares F test. For (L) and (M), data were pooled from 3 independent replicates. All plots show mean ± SEM.
    A 260, supplied by Agilent technologies, 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/a 260/product/Agilent technologies
    Average 86 stars, based on 1 article reviews
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    a 260 - by Bioz Stars, 2023-01
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    86
    PEQLAB a 260
    (A) Structure of <t>TDP43</t> RRM1 and RRM2, generated in FirstGlance in Jmol (PDB:4BS2), deposited by . TDP43 (blue) is shown in complex with UG-repeat RNA (green), highlighting the R151-D247 salt bridge (red). (B) TDP43 schematic depicting the RRM1-RRM2 salt bridge and F147-F149. (C) EMSA of increasing concentrations (12 fmol–4 pmol) of recombinant TDP43 variants mixed with labeled (UG) 12 (100 pM). (D, F, and H) Increasing concentrations (0.01–25 μM) of TDP43(WT) (D), TDP43(R151A) (F), and TDP43(D247A) (H) were added to labeled (UG) 12 (100 pM). (E, G, and I) K d and h were calculated from 3 independent replicates for TDP43(WT) (E), TDP43(R151A) (G), and TDP43(D247A) (I). (J and K) EMSA of TDP43 variants at increasing protein concentrations (0.5–16 pmol) incubated with 100 pM of labeled (GC) 12 (J) or (AT) 12 (K). (L and M) RNA IP of EGFP-tagged TDP43(WT), TDP43(F147L, F149L), TDP43(R151A), or TDP43(D247A) expressed in HEK293T cells. The abundance of bound TARDBP (L) or MALAT1 (M) transcripts was measured by qRT-PCR. ****p < 0.0001, one-way ANOVA with Dunnett’s test. Arrowheads in (C), (D), (F), (H), (J), and (K) indicate protein-DNA complexes, while arrows point to unbound oligonucleotides. For (E), (G), and (I) K d and h were determined by nonlinear least-squares regression; #p < 0.0001 for comparison with TDP43(WT) by extra sum-of-squares F test. For (L) and (M), data were pooled from 3 independent replicates. All plots show mean ± SEM.
    A 260, supplied by PEQLAB, 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/a 260/product/PEQLAB
    Average 86 stars, based on 1 article reviews
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    a 260 - by Bioz Stars, 2023-01
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    Image Search Results


    (A) Structure of TDP43 RRM1 and RRM2, generated in FirstGlance in Jmol (PDB:4BS2), deposited by . TDP43 (blue) is shown in complex with UG-repeat RNA (green), highlighting the R151-D247 salt bridge (red). (B) TDP43 schematic depicting the RRM1-RRM2 salt bridge and F147-F149. (C) EMSA of increasing concentrations (12 fmol–4 pmol) of recombinant TDP43 variants mixed with labeled (UG) 12 (100 pM). (D, F, and H) Increasing concentrations (0.01–25 μM) of TDP43(WT) (D), TDP43(R151A) (F), and TDP43(D247A) (H) were added to labeled (UG) 12 (100 pM). (E, G, and I) K d and h were calculated from 3 independent replicates for TDP43(WT) (E), TDP43(R151A) (G), and TDP43(D247A) (I). (J and K) EMSA of TDP43 variants at increasing protein concentrations (0.5–16 pmol) incubated with 100 pM of labeled (GC) 12 (J) or (AT) 12 (K). (L and M) RNA IP of EGFP-tagged TDP43(WT), TDP43(F147L, F149L), TDP43(R151A), or TDP43(D247A) expressed in HEK293T cells. The abundance of bound TARDBP (L) or MALAT1 (M) transcripts was measured by qRT-PCR. ****p < 0.0001, one-way ANOVA with Dunnett’s test. Arrowheads in (C), (D), (F), (H), (J), and (K) indicate protein-DNA complexes, while arrows point to unbound oligonucleotides. For (E), (G), and (I) K d and h were determined by nonlinear least-squares regression; #p < 0.0001 for comparison with TDP43(WT) by extra sum-of-squares F test. For (L) and (M), data were pooled from 3 independent replicates. All plots show mean ± SEM.

    Journal: Cell reports

    Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

    doi: 10.1016/j.celrep.2019.03.093

    Figure Lengend Snippet: (A) Structure of TDP43 RRM1 and RRM2, generated in FirstGlance in Jmol (PDB:4BS2), deposited by . TDP43 (blue) is shown in complex with UG-repeat RNA (green), highlighting the R151-D247 salt bridge (red). (B) TDP43 schematic depicting the RRM1-RRM2 salt bridge and F147-F149. (C) EMSA of increasing concentrations (12 fmol–4 pmol) of recombinant TDP43 variants mixed with labeled (UG) 12 (100 pM). (D, F, and H) Increasing concentrations (0.01–25 μM) of TDP43(WT) (D), TDP43(R151A) (F), and TDP43(D247A) (H) were added to labeled (UG) 12 (100 pM). (E, G, and I) K d and h were calculated from 3 independent replicates for TDP43(WT) (E), TDP43(R151A) (G), and TDP43(D247A) (I). (J and K) EMSA of TDP43 variants at increasing protein concentrations (0.5–16 pmol) incubated with 100 pM of labeled (GC) 12 (J) or (AT) 12 (K). (L and M) RNA IP of EGFP-tagged TDP43(WT), TDP43(F147L, F149L), TDP43(R151A), or TDP43(D247A) expressed in HEK293T cells. The abundance of bound TARDBP (L) or MALAT1 (M) transcripts was measured by qRT-PCR. ****p < 0.0001, one-way ANOVA with Dunnett’s test. Arrowheads in (C), (D), (F), (H), (J), and (K) indicate protein-DNA complexes, while arrows point to unbound oligonucleotides. For (E), (G), and (I) K d and h were determined by nonlinear least-squares regression; #p < 0.0001 for comparison with TDP43(WT) by extra sum-of-squares F test. For (L) and (M), data were pooled from 3 independent replicates. All plots show mean ± SEM.

    Article Snippet: Rabbit polyclonal anti-TDP43 , Cell Signaling Technologies , Cat#3449; RRID: AB_2200511.

    Techniques: Generated, Recombinant, Labeling, Incubation, Quantitative RT-PCR

    (A) Automated fluorescence microscopy. Individual neurons were programmatically identified (green outlines) and given unique identifiers (numbers). Cell death (red outlines) was determined by loss of fluorescence, rounding of the soma, or elimination of neuritic processes. Scale bar: 20 μm. (B) Overexpressing TDP43(WT)-EGFP significantly increases the cumulative risk of death compared to EGFP (HR = 5.04, p < 2 × 10 −16 ). Mutations that disrupt the salt bridge (R151A or D247A) or eliminate RNA binding (F147L, F149L) significantly reduce the risk of death compared to neurons expressing TDP43(WT) (HR = 0.25, 0.32, and 0.27, respectively; p < 2 × 10 −16 for all comparisons). (C) Deletion of RRM2 or both RRM1-RRM2 significantly improves survival compared to TDP43(WT) (HR = 0.51 and 0.31, respectively, p < 2 × 10 −16 for both comparisons). RRM1 deletion reduces the risk of death compared to TDP43(WT) (HR = 0.26, p < 2 × 10 −16 ) and EGFP alone (HR = 0.7, p < 2 × 10 −16 ). (D) Expression of TDP43(M337V)-EGFP significantly increases the risk of death (HR = 4.26, p < 2 × 10 −16 ) compared to EGFP. The F147L-F149L, R151A, and D247A mutations significantly reduce the toxicity of TDP43(M337V)-EGFP (HR = 0.34, 0.26, and 0.39, respectively; p < 2 × 10 −16 for all comparisons). n, number of neurons. ***p < 2 × 10 −16 ; Cox proportional hazards. Survival analyses were pooled from 3 independent experiments, with 8 wells per condition for each replicate.

    Journal: Cell reports

    Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

    doi: 10.1016/j.celrep.2019.03.093

    Figure Lengend Snippet: (A) Automated fluorescence microscopy. Individual neurons were programmatically identified (green outlines) and given unique identifiers (numbers). Cell death (red outlines) was determined by loss of fluorescence, rounding of the soma, or elimination of neuritic processes. Scale bar: 20 μm. (B) Overexpressing TDP43(WT)-EGFP significantly increases the cumulative risk of death compared to EGFP (HR = 5.04, p < 2 × 10 −16 ). Mutations that disrupt the salt bridge (R151A or D247A) or eliminate RNA binding (F147L, F149L) significantly reduce the risk of death compared to neurons expressing TDP43(WT) (HR = 0.25, 0.32, and 0.27, respectively; p < 2 × 10 −16 for all comparisons). (C) Deletion of RRM2 or both RRM1-RRM2 significantly improves survival compared to TDP43(WT) (HR = 0.51 and 0.31, respectively, p < 2 × 10 −16 for both comparisons). RRM1 deletion reduces the risk of death compared to TDP43(WT) (HR = 0.26, p < 2 × 10 −16 ) and EGFP alone (HR = 0.7, p < 2 × 10 −16 ). (D) Expression of TDP43(M337V)-EGFP significantly increases the risk of death (HR = 4.26, p < 2 × 10 −16 ) compared to EGFP. The F147L-F149L, R151A, and D247A mutations significantly reduce the toxicity of TDP43(M337V)-EGFP (HR = 0.34, 0.26, and 0.39, respectively; p < 2 × 10 −16 for all comparisons). n, number of neurons. ***p < 2 × 10 −16 ; Cox proportional hazards. Survival analyses were pooled from 3 independent experiments, with 8 wells per condition for each replicate.

    Article Snippet: Rabbit polyclonal anti-TDP43 , Cell Signaling Technologies , Cat#3449; RRID: AB_2200511.

    Techniques: Fluorescence, Microscopy, RNA Binding Assay, Expressing

    (A) Optical pulse labeling (OPL) of primary cortical neurons. Neurons transfected with EGFP and TDP43-Dendra2 variants were pulsed with 405 nm of light and imaged by automated microscopy. Scale bar: 50 μm. (B) The R151A, D247A, and F147L-F149L mutations significantly reduce the half-life of TDP43-Dendra2 ****p < 0.01, 1-way ANOVA with Dunnett’s test. (C) Cumulative frequency plot of protein half-life data. n, number of neurons. (D) Six hours before OPL, cells were treated with either DMSO or 500 nM MG132. ****p < 0.01, 1-way ANOVA with Tukey’s test. For (B), data pooled from 4 biological replicates. For (D), data accumulated from 400–2,000 cells per condition, over 3 biological replicates. Plots in (B) and (D) show median (horizontal line), interquartile range (box), and maximum/minimum (vertical lines).

    Journal: Cell reports

    Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

    doi: 10.1016/j.celrep.2019.03.093

    Figure Lengend Snippet: (A) Optical pulse labeling (OPL) of primary cortical neurons. Neurons transfected with EGFP and TDP43-Dendra2 variants were pulsed with 405 nm of light and imaged by automated microscopy. Scale bar: 50 μm. (B) The R151A, D247A, and F147L-F149L mutations significantly reduce the half-life of TDP43-Dendra2 ****p < 0.01, 1-way ANOVA with Dunnett’s test. (C) Cumulative frequency plot of protein half-life data. n, number of neurons. (D) Six hours before OPL, cells were treated with either DMSO or 500 nM MG132. ****p < 0.01, 1-way ANOVA with Tukey’s test. For (B), data pooled from 4 biological replicates. For (D), data accumulated from 400–2,000 cells per condition, over 3 biological replicates. Plots in (B) and (D) show median (horizontal line), interquartile range (box), and maximum/minimum (vertical lines).

    Article Snippet: Rabbit polyclonal anti-TDP43 , Cell Signaling Technologies , Cat#3449; RRID: AB_2200511.

    Techniques: Labeling, Transfection, Microscopy

    (A) Schematic of TDP43 hybrid constructs. (B) TDP43(PUM2)-EGFP expression significantly increased the cumulative risk of death compared to EGFP (HR = 2.54, p < 2 × 10 −16 ). Expression of the PUM2 RNA binding domain (RBD) marginally elevated the risk of death compared to EGFP (HR =1.19, p = 3.78 × 10 −5 ). RNA binding-deficient versions of the PUM2 RBD (mPUM2) significantly reduced the risk of death compared to TDP43(PUM2)-EGFP (HR = 0.45, p < 2 × 10 −16 ). (C) Images of neurons overexpressing mApple (red fluorescent protein [RFP]) and EGFP-tagged TDP43(WT), TDP43(PUM2), or TDP43(mPUM2). Scale bar: 5 μm. (D) Expression of TDP43(PP7) reduced the risk of death in comparison to TDP43(WT) (HR = 0.73, p < 2 × 10 −16 ) but remained significantly toxic compared to EGFP (HR = 2.05, p < 2 × 10 −16 ). (E) Boxplot of TDP43-Dendra2 variant half-life, determined by OPL. (F) Median TDP43-Dendra2 half-life decreased from 51.9 to 19.8 h with the addition of the destabilizing CL degron. (G) Both TDP43(WT)-Dendra2 and TDP43(WT)-Dendra2-CL significantly elevated the risk of death compared to Dendra2 (HR = 1.82 and 1.18, p < 2 × 10 −16 and p = 1.23 × 10 −10 , respectively). TDP43(WT)-Dendra2-CL was significantly less toxic compared to TDP43(WT)-Dendra2 (HR = 0.64, p < 2 × 10 −16 ). Data pooled from 3 independent replicates for (B) and (D) and 4 replicates for (E), (F), and (G). For (B) and (D)–(G), n, number of neurons; ***p < 2 × 10 −16 ; Cox proportional hazards. In (E), ***p < 0.05, 1-way ANOVA with Dunnett’s test. In (F), ****p < 0.0001, unpaired t test. Plots in (E) and (F) show median (horizontal line), interquartile range (box), and maximum/minimum (vertical lines).

    Journal: Cell reports

    Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

    doi: 10.1016/j.celrep.2019.03.093

    Figure Lengend Snippet: (A) Schematic of TDP43 hybrid constructs. (B) TDP43(PUM2)-EGFP expression significantly increased the cumulative risk of death compared to EGFP (HR = 2.54, p < 2 × 10 −16 ). Expression of the PUM2 RNA binding domain (RBD) marginally elevated the risk of death compared to EGFP (HR =1.19, p = 3.78 × 10 −5 ). RNA binding-deficient versions of the PUM2 RBD (mPUM2) significantly reduced the risk of death compared to TDP43(PUM2)-EGFP (HR = 0.45, p < 2 × 10 −16 ). (C) Images of neurons overexpressing mApple (red fluorescent protein [RFP]) and EGFP-tagged TDP43(WT), TDP43(PUM2), or TDP43(mPUM2). Scale bar: 5 μm. (D) Expression of TDP43(PP7) reduced the risk of death in comparison to TDP43(WT) (HR = 0.73, p < 2 × 10 −16 ) but remained significantly toxic compared to EGFP (HR = 2.05, p < 2 × 10 −16 ). (E) Boxplot of TDP43-Dendra2 variant half-life, determined by OPL. (F) Median TDP43-Dendra2 half-life decreased from 51.9 to 19.8 h with the addition of the destabilizing CL degron. (G) Both TDP43(WT)-Dendra2 and TDP43(WT)-Dendra2-CL significantly elevated the risk of death compared to Dendra2 (HR = 1.82 and 1.18, p < 2 × 10 −16 and p = 1.23 × 10 −10 , respectively). TDP43(WT)-Dendra2-CL was significantly less toxic compared to TDP43(WT)-Dendra2 (HR = 0.64, p < 2 × 10 −16 ). Data pooled from 3 independent replicates for (B) and (D) and 4 replicates for (E), (F), and (G). For (B) and (D)–(G), n, number of neurons; ***p < 2 × 10 −16 ; Cox proportional hazards. In (E), ***p < 0.05, 1-way ANOVA with Dunnett’s test. In (F), ****p < 0.0001, unpaired t test. Plots in (E) and (F) show median (horizontal line), interquartile range (box), and maximum/minimum (vertical lines).

    Article Snippet: Rabbit polyclonal anti-TDP43 , Cell Signaling Technologies , Cat#3449; RRID: AB_2200511.

    Techniques: Construct, Expressing, RNA Binding Assay, Variant Assay

    (A) Illustration of TDP43 variants tested in (B)–(F). (B and C) RNA IP of TARDBP (B) and MALAT1 (C) transcripts in HEK293T cells expressing EGFP-tagged TDP43 variants. (D) Significant decreases in protein half-life were observed for all TDP43 variants compared to TDP43(WT), as determined by OPL. (E) Cumulative frequency plot of data shown in (D). (F) By automated neuronal survival analysis, TDP43(WT)-EGFP was significantly toxic compared to EGFP (HR = 4.42, p < 2 × 10 −16 ). All TDP43 variants were less toxic than TDP43(WT)-EGFP (R151D, HR = 0.23; D247R, HR = 0.48; R151D-D247R, HR = 0.21; D247E, HR = 0.6; p < 2 × 10 −16 for all comparisons). Cells expressing either TDP43(D247R)-EGFP or TDP43(D247E)-EGFP demonstrated a significantly elevated risk of death compared to those expressing EGFP (HR = 2.2 and 2.76, respectively; p < 2 × 10 −16 for both comparisons). (G and H) While no relation was observed between half-life and toxicity for R151 variants (G), D247A variants displayed a linear relation between half-life and toxicity (H). For (B) and (C), data were pooled from 3 independent experiments, ****p < 0.05, Kruskal-Wallis with Dunn’s test. For (D), data were pooled from 4 replicates, ****p < 0.05, 1-way ANOVA with Dunnett’s test. For (F), data represent 3 replicates, ***p < 2 × 10 −16 , Cox proportional hazards. n, number of neurons. For (G) and(H), error bars represent 95% confidence intervals. In (H), r 2 calculated by nonlinear least-squares regression.

    Journal: Cell reports

    Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

    doi: 10.1016/j.celrep.2019.03.093

    Figure Lengend Snippet: (A) Illustration of TDP43 variants tested in (B)–(F). (B and C) RNA IP of TARDBP (B) and MALAT1 (C) transcripts in HEK293T cells expressing EGFP-tagged TDP43 variants. (D) Significant decreases in protein half-life were observed for all TDP43 variants compared to TDP43(WT), as determined by OPL. (E) Cumulative frequency plot of data shown in (D). (F) By automated neuronal survival analysis, TDP43(WT)-EGFP was significantly toxic compared to EGFP (HR = 4.42, p < 2 × 10 −16 ). All TDP43 variants were less toxic than TDP43(WT)-EGFP (R151D, HR = 0.23; D247R, HR = 0.48; R151D-D247R, HR = 0.21; D247E, HR = 0.6; p < 2 × 10 −16 for all comparisons). Cells expressing either TDP43(D247R)-EGFP or TDP43(D247E)-EGFP demonstrated a significantly elevated risk of death compared to those expressing EGFP (HR = 2.2 and 2.76, respectively; p < 2 × 10 −16 for both comparisons). (G and H) While no relation was observed between half-life and toxicity for R151 variants (G), D247A variants displayed a linear relation between half-life and toxicity (H). For (B) and (C), data were pooled from 3 independent experiments, ****p < 0.05, Kruskal-Wallis with Dunn’s test. For (D), data were pooled from 4 replicates, ****p < 0.05, 1-way ANOVA with Dunnett’s test. For (F), data represent 3 replicates, ***p < 2 × 10 −16 , Cox proportional hazards. n, number of neurons. For (G) and(H), error bars represent 95% confidence intervals. In (H), r 2 calculated by nonlinear least-squares regression.

    Article Snippet: Rabbit polyclonal anti-TDP43 , Cell Signaling Technologies , Cat#3449; RRID: AB_2200511.

    Techniques: Expressing

    (A) TDP43 schematic depicting variants used for RNA sequencing (RNA-seq). (B) Venn diagram illustrating differentially expressed genes (DEGs) for each condition, in comparison to EGFP alone. (C) TARDBP transcript levels were similar among all TDP43 variants, demonstrating equivalent overexpression in each case. (D) Venn diagram showing significant enrichment for functional pathways according to GO analysis. (E–G) Among downregulated mRNAs, transcripts involved in the ribosomal (E) and oxidative phosphorylation (F) pathways were highly enriched (FDR = 6.8 × 10 −5 and 0.03, respectively), with a trend toward enrichment for transcripts related to RNA transport (G, FDR = 0.222).

    Journal: Cell reports

    Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

    doi: 10.1016/j.celrep.2019.03.093

    Figure Lengend Snippet: (A) TDP43 schematic depicting variants used for RNA sequencing (RNA-seq). (B) Venn diagram illustrating differentially expressed genes (DEGs) for each condition, in comparison to EGFP alone. (C) TARDBP transcript levels were similar among all TDP43 variants, demonstrating equivalent overexpression in each case. (D) Venn diagram showing significant enrichment for functional pathways according to GO analysis. (E–G) Among downregulated mRNAs, transcripts involved in the ribosomal (E) and oxidative phosphorylation (F) pathways were highly enriched (FDR = 6.8 × 10 −5 and 0.03, respectively), with a trend toward enrichment for transcripts related to RNA transport (G, FDR = 0.222).

    Article Snippet: Rabbit polyclonal anti-TDP43 , Cell Signaling Technologies , Cat#3449; RRID: AB_2200511.

    Techniques: RNA Sequencing Assay, Over Expression, Functional Assay

    (A–D) A total of 9,339 differential splicing events involving 4,819 genes were noted upon TDP43(WT)-EGFP expression (A). Of these, 34.2% were also detected in cells expressing TDP43(F147L, F149L)-EGFP (B), 29.3% with TDP43(R151A)-EGFP (C), and 38.7% with TDP43(D247A)-EGFP (D). (E) A total of 3,968 transcripts (42.5%) were selectively spliced by TDP43(WT)-EGFP. (F) Heatmap of the 50 top repressed and 50 top included splicing events. See also . (G) Categorization of TDP43(WT)-specific splicing events. RI, intron retention; A3SS, alternative 3 ′ splice sites; A5SS, alternative 5 ′ splice sites; ALE, alternative last exon; AFE, alternative first exon; AS, alternative start; SE, skipping of 1 exon; S2E, skipping of 2 exons; MXE, mutually exclusive exons. (H and I) Representative examples of abnormal splicing in NDUFS6 (H) and ITPR3 (I) transcripts, with schematic diagrams of exons (E) and splice junctions (J) above heatmaps displaying the read density for each junction. Unannotated exons/junctions are shown in red.

    Journal: Cell reports

    Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

    doi: 10.1016/j.celrep.2019.03.093

    Figure Lengend Snippet: (A–D) A total of 9,339 differential splicing events involving 4,819 genes were noted upon TDP43(WT)-EGFP expression (A). Of these, 34.2% were also detected in cells expressing TDP43(F147L, F149L)-EGFP (B), 29.3% with TDP43(R151A)-EGFP (C), and 38.7% with TDP43(D247A)-EGFP (D). (E) A total of 3,968 transcripts (42.5%) were selectively spliced by TDP43(WT)-EGFP. (F) Heatmap of the 50 top repressed and 50 top included splicing events. See also . (G) Categorization of TDP43(WT)-specific splicing events. RI, intron retention; A3SS, alternative 3 ′ splice sites; A5SS, alternative 5 ′ splice sites; ALE, alternative last exon; AFE, alternative first exon; AS, alternative start; SE, skipping of 1 exon; S2E, skipping of 2 exons; MXE, mutually exclusive exons. (H and I) Representative examples of abnormal splicing in NDUFS6 (H) and ITPR3 (I) transcripts, with schematic diagrams of exons (E) and splice junctions (J) above heatmaps displaying the read density for each junction. Unannotated exons/junctions are shown in red.

    Article Snippet: Rabbit polyclonal anti-TDP43 , Cell Signaling Technologies , Cat#3449; RRID: AB_2200511.

    Techniques: Expressing

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

    doi: 10.1016/j.celrep.2019.03.093

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Rabbit polyclonal anti-TDP43 , Cell Signaling Technologies , Cat#3449; RRID: AB_2200511.

    Techniques: Recombinant, Clone Assay, Mutagenesis, Labeling, Plasmid Preparation, Software, Fractionation