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Image Search Results
Journal: The Journal of Biological Chemistry
Article Title: High-throughput screening yields several small-molecule inhibitors of repeat-associated non-AUG translation
doi: 10.1074/jbc.RA119.009951
Figure Lengend Snippet: Small molecules inhibit CGG RAN translation in multiple reading frames. A, structures of compounds used in B–H. B–F, compounds identified from screen as RAN translation inhibitors were independently reassessed for their activity in RRL in vitro translation assays. The indicated compounds were added at increasing doses to 30% RRL reactions with AUG, +1CGGx100, or +2CGGx100-NLuc reporter mRNAs, and luminescence was measured relative to vehicle (DMSO) treatment. G and H, candidate compounds protoporphyrin IX and TMPyP4 were added at increasing concentrations to RRL reactions with AUG, +1CGGx100, or +2CGGx100-NLuc reporter mRNAs. Luminescence was measured relative to vehicle (DMSO) treatment. For all graphs, n = 3; error bars, S.D. *, p < 0.05, two-way ANOVA with Dunnett's multiple-comparison test.
Article Snippet: Independent validation of hits in RRL system The following compounds were purchased through Sigma Marketsite: cephalothin sodium (Sigma–Aldrich, C4520), rufloxacin HCl (Vitas-M Laboratory, Ltd., STK711124), rofecoxib (Sigma–Aldrich, MFCD00935806), alfuzosin HCl (Sigma–Aldrich, A0232), olanzapine (Sigma–Aldrich, O1141),
Techniques: Activity Assay, In Vitro, Comparison
Journal: The Journal of Biological Chemistry
Article Title: High-throughput screening yields several small-molecule inhibitors of repeat-associated non-AUG translation
doi: 10.1074/jbc.RA119.009951
Figure Lengend Snippet: Small molecules inhibit C9RAN translation in all three reading frames. BIX01294 (A), anthralin (B), and protoporphyrin IX (C) were added at increasing concentrations to 30% RRL in vitro translation reactions with AUG-NLuc or GGGGCCx70 reporter mRNAs for all three reading frames, and luminescence was measured relative to vehicle (DMSO) control. D–F, selective inhibition of RAN translation of GGGGCCx70 and CGGx100 NLuc reporter mRNAs, relative to AUG-NLuc control, was confirmed by Western blotting against a C-terminal FLAG tag for the glycine-alanine (GA) and FMRPolyG (+1CGG) products, following incubation with BIX01294, anthralin, or protoporphyrin IX at the indicated doses. GAPDH was used as a loading control. For A–C, all graphs represent mean of n = 3; error bars, S.D. *, p < 0.05, two-way ANOVA with Dunnett's multiple-comparison test. M, mock; GA, glycine-alanine; GP, glycine-proline; GR, glycine-arginine.
Article Snippet: Independent validation of hits in RRL system The following compounds were purchased through Sigma Marketsite: cephalothin sodium (Sigma–Aldrich, C4520), rufloxacin HCl (Vitas-M Laboratory, Ltd., STK711124), rofecoxib (Sigma–Aldrich, MFCD00935806), alfuzosin HCl (Sigma–Aldrich, A0232), olanzapine (Sigma–Aldrich, O1141),
Techniques: In Vitro, Inhibition, Western Blot, FLAG-tag, Incubation, Comparison
Journal: The Journal of Biological Chemistry
Article Title: High-throughput screening yields several small-molecule inhibitors of repeat-associated non-AUG translation
doi: 10.1074/jbc.RA119.009951
Figure Lengend Snippet: RAN translation inhibitors have varying effects on AUG-initiated repeat translation. A, schematic of NLuc reporter mRNAs used in 30% RRL in vitro translation assays, illustrating the position of the AUG start codons inserted upstream of the repeats, to drive translation in the indicated reading frames. The effect of anthralin (B), TMPyP4 (C), protoporphyrin IX (D), BIX01294 (E), and CP-31398 (F) on +1 and +2CGGx100 RAN translation, compared with AUG-initiated CGGx100 translation, measured in the presence of increasing concentrations of each small molecule. For B–F, values are relative to DMSO controls, and graphs represent the mean of n = 3. Error bars, S.D. *, p < 0.05, two-way ANOVA with Dunnett's multiple-comparison test. 3xF, 3xFLAG tag.
Article Snippet: Independent validation of hits in RRL system The following compounds were purchased through Sigma Marketsite: cephalothin sodium (Sigma–Aldrich, C4520), rufloxacin HCl (Vitas-M Laboratory, Ltd., STK711124), rofecoxib (Sigma–Aldrich, MFCD00935806), alfuzosin HCl (Sigma–Aldrich, A0232), olanzapine (Sigma–Aldrich, O1141),
Techniques: In Vitro, Comparison