uncropped gel images  (Mendeley Ltd)

Journal: iScience

Article Title: TM4SF5-mediated abnormal food-intake behavior and apelin expression facilitate non-alcoholic fatty liver disease features

doi: 10.1016/j.isci.2023.107625

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Article Snippet: Uncropped raw immunoblot gel images , Mendeley Data , https://data.mendeley.com/datasets/y9wwtjy74h/1.

Techniques: Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Plasmid Preparation, Bioassay, Sequencing, Western Blot, Transgenic Assay, shRNA, Software

Journal: PLoS ONE

Article Title: Image-based screen capturing misfolding status of Niemann-Pick type C1 identifies potential candidates for chaperone drugs

doi: 10.1371/journal.pone.0243746

Figure Lengend Snippet: (A) High-resolution clear native PAGE (fluorescence-detectable native PAGE) analysis of NPC1-GFP from cells treated as indicated. Cells stably expressing the indicated NPC1-GFP were treated with vehicle, mo56HC (3 μM), itraconazole (3 μM), or lapatinib (10 μM) for 21 h, and DDM-solubilized lysates were subjected to native-PAGE analysis after normalization with respect to total protein concentration. The native-PAGE gel was imaged for GFP fluorescence (upper panel), and total protein was also visualized by TCE staining (lower panel). (B) Ubiquitination status of NPC1-GFPs upon treatment with itraconazole or lapatinib. Cells stably expressing the indicated NPC1-GFP were treated as shown for 18 h followed by treatment with CB5083 (3 μM) for 6 h. The ubiquitination status was first probed with anti-Ub antibody after immunoprecipitation of the FLAG-NPC1-GFP, and then the amount of the FLAG-NPC1-GFP was re-probed with anti-FLAG antibody. The extent of ubiquitination was quantified and normalized to the amount of detected FLAG-NPC1-GFP. The raw, uncropped blot images are available from Mendeley Data repository ( http://dx.doi.org/10.17632/jr23ccpp46.3 ). (C) Quantification of the high-resolution clear native PAGE analysis from three independent experiments. The filled circles, error bars, and open circles represent mean, SEM, and raw data points from three independent experiments. Statistical significance was assessed by ANOVA and Tukey-Kramer multiple comparison test (***, p<0.001). The raw, uncropped gel images are available from Mendeley Data repository ( http://dx.doi.org/10.17632/jr23ccpp46.3 ). (D) Quantification of the ubiquitination assay data collected from three independent experiments. Data was normalized with the ubiquitination level of WT for each experiment. The filled circles, error bars, and open circles represent mean, SEM, and raw data points. Statistical significance of the treatment was assessed similarly as in (C). *, p<0.05; p<0.1; ns, not significant (p = 0.24).

Article Snippet: Other relevant data, including R scripts for analysis, raw uncropped gel images, and quantitative data used for preparing the figures are available from Mendeley Data ( https://doi.org/10.17632/jr23ccpp46 ).

Techniques: Clear Native PAGE, Fluorescence, Stable Transfection, Expressing, Protein Concentration, Staining, Ubiquitin Proteomics, Immunoprecipitation, Comparison

Journal: PLoS ONE

Article Title: Image-based screen capturing misfolding status of Niemann-Pick type C1 identifies potential candidates for chaperone drugs

doi: 10.1371/journal.pone.0243746

Figure Lengend Snippet: (A) Structures of photoaffinity probes and pull-down probes for itraconazole and lapatinib. (B) Chaperone activity of the probes on NPC1-I1061T mutant protein, examined by image-based chaperone assay. (C) Photoaffinity labeling of FLAG-tagged NPC1-I1061T-GFP or NPC1-P691S/I1061T-GFP with itraAZY probe. Membrane fractions were prepared from the cells stably expressing the indicated NPC1-GFP, and photo-crosslinking was performed in the presence or absence of the indicated competitors. After conjugation of biotin to the alkyne via click chemistry, the NPC1 was immunoprecipitated and the presence of biotin was probed with streptavidin-HRP conjugate, followed by re-probing with anti-FLAG antibody. The raw, uncropped blot images are available from Mendeley Data repository ( http://dx.doi.org/10.17632/jr23ccpp46.3 ). (D) Quantification of the labeling. The extent of labeling was quantified and normalized with respect to the amount of the FLAG-NPC1-GFPs. Data is from independent experiments (n = 4 for I1061T and n = 3 for P691S/I1061T), and expressed as relative labeling respect to I1061T labeled with itraAZY. The filled circles, open circles, and error bars represent mean, raw data, and SEM, respectively. Statistical significance was assessed by the exact Wilcoxon rank sum test with correction of multiple comparisons by Benjamini-Hochberg method (*, p = 0.05; ns, not significant).

Article Snippet: Other relevant data, including R scripts for analysis, raw uncropped gel images, and quantitative data used for preparing the figures are available from Mendeley Data ( https://doi.org/10.17632/jr23ccpp46 ).

Techniques: Activity Assay, Mutagenesis, Labeling, Membrane, Stable Transfection, Expressing, Conjugation Assay, Immunoprecipitation