inactive sirt1 h363y mutant (Addgene inc)
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inactive sirt1 h363y mutant
Inactive Sirt1 H363y Mutant, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mutant/Flag-SIRT1+H363Y+(Plasmid+%231792)/pmc13684318-319-13-19
Average 93 stars, based on 79 article reviews
Inactive Sirt1 H363y Mutant, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mutant/Flag-SIRT1+H363Y+(Plasmid+%231792)/pmc13684318-319-13-19
Average 93 stars, based on 79 article reviews
inactive sirt1 h363y mutant - by Bioz Stars,
2026-11
93/100 stars
Images
1) Product Images from "A Cilia–Centered Nicotinamide Adenine Dinucleotide ( NAD + ) Signaling Network Regulates Hypothalamic Lysosomal Homeostasis and Metabolic Aging"
Article Title: A Cilia–Centered Nicotinamide Adenine Dinucleotide ( NAD + ) Signaling Network Regulates Hypothalamic Lysosomal Homeostasis and Metabolic Aging
Journal: Aging Cell
doi: 10.1111/acel.70745
Figure Legend Snippet: SIRT1 mediates the regulatory effect of NAD + on primary cilia. (a) SIRT1 activator resveratrol (20 μM) treatment elongates cilia length in N1 cells ( n = 8 wells per group). AC3: Adenylyl cyclase 3. (b) Effects of overexpression (OE) of wild–type SIRT1 and its catalytically inactive H363Y mutant on ciliary length in N1 cells ( n = 5 wells per group). (c) Effect of treatment with the SIRT1 inhibitor EX527 (100 μm for 16 h) on NR (100 μM)‐induced cilia growth ( n = 6 wells per group). (d) Sirt1 knockdown with small inhibitory RNA ( siSirt1 ) prevents NMN (100 μM)–induced cilia elongation ( n = 3 wells per group). (e) Effect of SIRT1 OE on mRNA expression levels of multiple cilia‐related genes ( n = 6 wells per group). (f) Sirt1 knockdown inhibits NMN‐induced upregulation of cilia‐related genes ( n = 6 wells per group). (g, h) SIRT1 and RFX1 cooperatively regulate the promoter activity of the Ift88 gene ( n = 3 wells per group). (i, j) Molecular interaction between SIRT1 and RFX1, and SIRT1‐mediated deacetylation of RFX1, demonstrated by sequential immunoprecipitation (IP) and immunoblotting (IB) in N1 cells with or without SIRT1 OE. Ac‐K, acetylated lysine. (k) Effect of autophagy inhibition by Atg5 knockdown or 3‐MA treatment on SIRT1‐induced changes in primary cilia ( n = 5). Data were obtained from three independent experiments and are presented as the mean ± SEM. Statistical analyses were performed using two‐sided unpaired Student's t‐ test (a, e), one‐way ANOVA (b, c, f, g, h, k), or two‐way (d) ANOVA with Tukey's post hoc test. n.s., not significant. Scale bars, 20 μm.
Techniques Used: Over Expression, Mutagenesis, Knockdown, Expressing, Activity Assay, Immunoprecipitation, Western Blot, Inhibition
Figure Legend Snippet: Primary cilia are essential for NAD + ‐mediated regulation of lysosomal acidity. (a) Effects of NAD + elevation induced by NAD or its precursors (NMN, NR) treatment on lysosomal acidity, assessed by LysoSensor fluorescence intensity ( n = 6 wells per group). Scale bars, 20 μm. (b) Effect of overexpression of wild‐type SIRT1 and a catalytically inactive SIRT1 mutant ( H363Y ) on lysosomal acidity ( n = 5 wells per group). (c, d) Effects of NAD + depletion induced by FK866 treatment or Nampt knockdown on lysosomal acidity ( n = 6 wells per group). Scale bars, 20 μm. (e) Restorative effects of NMN co‐treatment on the FK866‐induced lysosomal deacidification in N1 cells transfected with control siRNA ( siControl ), Ift88 siRNA ( siIft88 ), or Ift20 siRNA ( siIft20 ) ( n = 4 wells per group). Scale bars, 20 μm. (f) Effects of NAD + depletion (FK866) and repletion (FK866 + NMN) on lysosomal cathepsin B activity, assessed by the Magic Red assay, in N1 cells with or without ciliary dysgenesis ( n = 6 wells per group). Data were obtained from three independent experiments and are presented as the mean ± SEM. Statistical analyses were performed using two‐sided unpaired Student's t ‐test (c, d), one‐way ANOVA (a, b), or two‐way ANOVA (e, f), followed by Tukey's post hoc test. n.s., not significant.
Techniques Used: Fluorescence, Over Expression, Mutagenesis, Knockdown, Transfection, Control, Activity Assay
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