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Fig. 4 | Sestrin is important for maintenance of ISCs in response to DR. a,b, Gut stem cell activity of wDah and Sesn3F6 flies under DR. a, Representative images from 20-day-old flies. Actively dividing stem cells are indicated by arrows. b, Quantification of <t>pH3-positive</t> cells in Sesn3F6 females under DR. n = 25 guts (wDah, Sesn3F6, 2.0×; Sesn3F6, 1.0×), n = 24 guts (wDah, 1.0×). c,d, Gut turnover rates on Sestrin knockdown (KD). c, Representative images after 7 d induction. d, Quantification of gut turnover rates in Sestrin knockdown flies under DR. n = 14 guts (wDah, 2.0×, 1.0×), n = 16 guts (Sesn KD, 2.0×), n = 17 guts (Sesn KD, 1.0×). e,f, Gut stem cell activity of SesnR407A mutant flies under DR conditions. e, Representative images from 20-day-old flies. f, Quantification of pH3-positive cells in SesnR407A mutants under DR. n = 25 guts (Sesnwt, 2.0×, 1.0×), n = 24 guts (SesnR407A, 2.0×, 1.0×). g,h, Gut turnover rates in SesnR407A mutant flies under DR. g, Representative images after 7 d induction. h, Quantification of gut turnover rates in SesnR407A mutants under DR. n = 14 guts (Sesnwt, 2.0×; SesnR407A, 1.0×), n = 15 guts (Sesnwt, 1.0×; SesnR407A, 2.0×). i,j, Quantification of gut turnover rates in SesnR407A mutants on addition of all amino acids (i) and addition of methionine and BCAAs (MBC) (j). n = 13 guts (i), n = 16 guts (j). Scale bars, 50 μm. Median, 25th and 75th percentiles, and Tukey whiskers are indicated in box-and-whisker plots (b,d,f,h–j). Outliers are shown as open circles. Interaction between diet and genotype was significant in b,d,f and h–j: two-way ANOVA, P = 0.02 (b), P = 0.01 (d), P = 0.02 (f), P = 0.01 (h), P < 0.0001 (i), P = 0.047 (j). Statistics in b,d,f and h–j: two-way ANOVA followed by Bonferroni’s post-hoc test, P values were adjusted for multiple comparisons.

Ph3 S10 Rabbit Polyclonal, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 2931 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Sestrin is a key regulator of stem cell function and lifespan in response to dietary amino acids."

Article Title: Sestrin is a key regulator of stem cell function and lifespan in response to dietary amino acids.

Journal: Nature aging

doi: 10.1038/s43587-020-00001-7

Figure Legend Snippet: Fig. 4 | Sestrin is important for maintenance of ISCs in response to DR. a,b, Gut stem cell activity of wDah and Sesn3F6 flies under DR. a, Representative images from 20-day-old flies. Actively dividing stem cells are indicated by arrows. b, Quantification of pH3-positive cells in Sesn3F6 females under DR. n = 25 guts (wDah, Sesn3F6, 2.0×; Sesn3F6, 1.0×), n = 24 guts (wDah, 1.0×). c,d, Gut turnover rates on Sestrin knockdown (KD). c, Representative images after 7 d induction. d, Quantification of gut turnover rates in Sestrin knockdown flies under DR. n = 14 guts (wDah, 2.0×, 1.0×), n = 16 guts (Sesn KD, 2.0×), n = 17 guts (Sesn KD, 1.0×). e,f, Gut stem cell activity of SesnR407A mutant flies under DR conditions. e, Representative images from 20-day-old flies. f, Quantification of pH3-positive cells in SesnR407A mutants under DR. n = 25 guts (Sesnwt, 2.0×, 1.0×), n = 24 guts (SesnR407A, 2.0×, 1.0×). g,h, Gut turnover rates in SesnR407A mutant flies under DR. g, Representative images after 7 d induction. h, Quantification of gut turnover rates in SesnR407A mutants under DR. n = 14 guts (Sesnwt, 2.0×; SesnR407A, 1.0×), n = 15 guts (Sesnwt, 1.0×; SesnR407A, 2.0×). i,j, Quantification of gut turnover rates in SesnR407A mutants on addition of all amino acids (i) and addition of methionine and BCAAs (MBC) (j). n = 13 guts (i), n = 16 guts (j). Scale bars, 50 μm. Median, 25th and 75th percentiles, and Tukey whiskers are indicated in box-and-whisker plots (b,d,f,h–j). Outliers are shown as open circles. Interaction between diet and genotype was significant in b,d,f and h–j: two-way ANOVA, P = 0.02 (b), P = 0.01 (d), P = 0.02 (f), P = 0.01 (h), P < 0.0001 (i), P = 0.047 (j). Statistics in b,d,f and h–j: two-way ANOVA followed by Bonferroni’s post-hoc test, P values were adjusted for multiple comparisons.

Techniques Used: Activity Assay, Knockdown, Mutagenesis, Whisker Assay

Fig. 5 | Sestrin overexpression in gut stem cells improves gut homeostasis. a,b, Ubiquitous overexpression of Sestrin reduced gut stem cell activity. a, Representative images from 30-day-old flies without (RU486−) or with (RU486+) induction. Actively dividing stem cells are indicated by arrows. Scale bar, 50 μm. b, Quantification of pH3-positive cells on days 10, 30 and 50. n = 20 guts (RU486−), n = 22 guts (RU486+), day 10; n = 23 guts (RU486−, RU486+), day 30; n = 24 guts (RU486−, RU486+), day 50. Generalized linear modelling analysis confirmed that Sestrin overexpression significantly reduced the increase in age-dependent stem cell activity (P = 0.012). c, Overexpression of Sestrin in enterocytes (5966GS) did not affect pH3-positive cell numbers. Day 30: n = 22 guts (RU486−), n = 21 guts (RU486+); day 46: n = 22 guts (RU486−), n = 20 guts (RU486+). d, In contrast, overexpression of Sestrin in gut stem cells (5961GS) significantly reduced stem cell activity. Day 30: n = 24 guts (RU486−), n = 25 guts (RU486+); day 46: n = 22 guts (RU486−), n = 23 guts (RU486+). e, Sestrin overexpression (OE) prolonged the G1 phase in gut stem cells. Fly-Fucci and Sestrin were driven by the esg-Gal4 driver. Sestrin overexpression significantly changed the cell cycle distribution of gut stem cells. n = 416 cells, wDah; n = 382 cells, Sesn OE. Chi-square test. f–h, Sestrin overexpression in gut stem cells decreased gut dysplasia (f,g) and the proportion of Smurf flies (h). f, Representative gut images from 50-day-old flies. DNA (DAPI) in blue. The gut epithelium is indicated by dashed lines. Scale bar, 20 μm. g, The difference in gut dysplasia was significant. n = 13 guts. h, Flies were 60 days old. n = 15 vials. Median, 25th and 75th percentiles, and Tukey whiskers are indicated in box-and-whisker plots (b–d,g,h). Outliers are shown as open circles. Statistics in b–d: two-tailed, Mann–Whitney test; statistics in g and h: unpaired, two-tailed t-test.

Figure Legend Snippet: Fig. 5 | Sestrin overexpression in gut stem cells improves gut homeostasis. a,b, Ubiquitous overexpression of Sestrin reduced gut stem cell activity. a, Representative images from 30-day-old flies without (RU486−) or with (RU486+) induction. Actively dividing stem cells are indicated by arrows. Scale bar, 50 μm. b, Quantification of pH3-positive cells on days 10, 30 and 50. n = 20 guts (RU486−), n = 22 guts (RU486+), day 10; n = 23 guts (RU486−, RU486+), day 30; n = 24 guts (RU486−, RU486+), day 50. Generalized linear modelling analysis confirmed that Sestrin overexpression significantly reduced the increase in age-dependent stem cell activity (P = 0.012). c, Overexpression of Sestrin in enterocytes (5966GS) did not affect pH3-positive cell numbers. Day 30: n = 22 guts (RU486−), n = 21 guts (RU486+); day 46: n = 22 guts (RU486−), n = 20 guts (RU486+). d, In contrast, overexpression of Sestrin in gut stem cells (5961GS) significantly reduced stem cell activity. Day 30: n = 24 guts (RU486−), n = 25 guts (RU486+); day 46: n = 22 guts (RU486−), n = 23 guts (RU486+). e, Sestrin overexpression (OE) prolonged the G1 phase in gut stem cells. Fly-Fucci and Sestrin were driven by the esg-Gal4 driver. Sestrin overexpression significantly changed the cell cycle distribution of gut stem cells. n = 416 cells, wDah; n = 382 cells, Sesn OE. Chi-square test. f–h, Sestrin overexpression in gut stem cells decreased gut dysplasia (f,g) and the proportion of Smurf flies (h). f, Representative gut images from 50-day-old flies. DNA (DAPI) in blue. The gut epithelium is indicated by dashed lines. Scale bar, 20 μm. g, The difference in gut dysplasia was significant. n = 13 guts. h, Flies were 60 days old. n = 15 vials. Median, 25th and 75th percentiles, and Tukey whiskers are indicated in box-and-whisker plots (b–d,g,h). Outliers are shown as open circles. Statistics in b–d: two-tailed, Mann–Whitney test; statistics in g and h: unpaired, two-tailed t-test.

Techniques Used: Over Expression, Activity Assay, Whisker Assay, Two Tailed Test, MANN-WHITNEY

Biomarker Discovery:

Article Title: Sestrin is a key regulator of stem cell function and lifespan in response to dietary amino acids
Article Snippet: .. Lee Anti pH3 (S10) rabbit polyclonal, Cell signaling, Cat# 9701 Anti Myc (9B11) mouse monoclonal, Cell signaling, Cat# 2276 Anti Flag M2 mouse monoclonal, Sigma, Cat# F1804 Anti HA rabbit polyclonal, Sigma, Cat# H6908 Anti pS6K (T398) rabbit polyclonal, Cell signaling, Cat# 9209 Anti S6K rabbit polyclonal, home-made Anti pAMPKα (T172) (40H9) rabbit monoclonal, Cell signaling, Cat# 2535 Anti pAKT (S505) rabbit polyclonal, Cell signaling, Cat# 4054 Anti AKT rabbit polyclonal, Cell signaling, Cat# 9272 Anti Tubulin (DM1A) mouse monoclonal, Sigma, Cat# T9026 Goat anti-rabbit IgG IRDye 680RD, LI-COR, Cat# 926-68071 Goat anti-mouse IgG IRDye 800CW, LI-COR, Cat# 926-32210 Goat anti-rabbit IgG (H+L), Alexa Fluor 594, Thermo Fisher Scientific, Cat# A11012 Validation Anti Sestrin rabbit polyclonal, home-made, gift from J.H. ..

Article Title: Sestrin is a key regulator of stem cell function and lifespan in response to dietary amino acids.
Article Snippet: .. Lee Anti pH3 (S10) rabbit polyclonal, Cell signaling, Cat# 9701 Anti Myc (9B11) mouse monoclonal, Cell signaling, Cat# 2276 Anti Flag M2 mouse monoclonal, Sigma, Cat# F1804 Anti HA rabbit polyclonal, Sigma, Cat# H6908 Anti pS6K (T398) rabbit polyclonal, Cell signaling, Cat# 9209 Anti S6K rabbit polyclonal, home-made Anti pAMPKα (T172) (40H9) rabbit monoclonal, Cell signaling, Cat# 2535 Anti pAKT (S505) rabbit polyclonal, Cell signaling, Cat# 4054 Anti AKT rabbit polyclonal, Cell signaling, Cat# 9272 Anti Tubulin (DM1A) mouse monoclonal, Sigma, Cat# T9026 Goat anti-rabbit IgG IRDye 680RD, LI-COR, Cat# 926-68071 Goat anti-mouse IgG IRDye 800CW, LI-COR, Cat# 926-32210 Goat anti-rabbit IgG (H+L), Alexa Fluor 594, Thermo Fisher Scientific, Cat# A11012 Validation Anti Sestrin rabbit polyclonal, home-made, gift from J.H. ..

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Journal: Nature aging

Article Title: Sestrin is a key regulator of stem cell function and lifespan in response to dietary amino acids.

doi: 10.1038/s43587-020-00001-7

Figure Lengend Snippet: Fig. 4 | Sestrin is important for maintenance of ISCs in response to DR. a,b, Gut stem cell activity of wDah and Sesn3F6 flies under DR. a, Representative images from 20-day-old flies. Actively dividing stem cells are indicated by arrows. b, Quantification of pH3-positive cells in Sesn3F6 females under DR. n = 25 guts (wDah, Sesn3F6, 2.0×; Sesn3F6, 1.0×), n = 24 guts (wDah, 1.0×). c,d, Gut turnover rates on Sestrin knockdown (KD). c, Representative images after 7 d induction. d, Quantification of gut turnover rates in Sestrin knockdown flies under DR. n = 14 guts (wDah, 2.0×, 1.0×), n = 16 guts (Sesn KD, 2.0×), n = 17 guts (Sesn KD, 1.0×). e,f, Gut stem cell activity of SesnR407A mutant flies under DR conditions. e, Representative images from 20-day-old flies. f, Quantification of pH3-positive cells in SesnR407A mutants under DR. n = 25 guts (Sesnwt, 2.0×, 1.0×), n = 24 guts (SesnR407A, 2.0×, 1.0×). g,h, Gut turnover rates in SesnR407A mutant flies under DR. g, Representative images after 7 d induction. h, Quantification of gut turnover rates in SesnR407A mutants under DR. n = 14 guts (Sesnwt, 2.0×; SesnR407A, 1.0×), n = 15 guts (Sesnwt, 1.0×; SesnR407A, 2.0×). i,j, Quantification of gut turnover rates in SesnR407A mutants on addition of all amino acids (i) and addition of methionine and BCAAs (MBC) (j). n = 13 guts (i), n = 16 guts (j). Scale bars, 50 μm. Median, 25th and 75th percentiles, and Tukey whiskers are indicated in box-and-whisker plots (b,d,f,h–j). Outliers are shown as open circles. Interaction between diet and genotype was significant in b,d,f and h–j: two-way ANOVA, P = 0.02 (b), P = 0.01 (d), P = 0.02 (f), P = 0.01 (h), P < 0.0001 (i), P = 0.047 (j). Statistics in b,d,f and h–j: two-way ANOVA followed by Bonferroni’s post-hoc test, P values were adjusted for multiple comparisons.

Article Snippet: Science 327, 1223-1228 3 n atu re research | rep o rtin g su m m ary O c to b e r 2 0 1 8 Anti pH3 (S10) rabbit polyclonal, Cell signaling, Cat# 9701.

Techniques: Activity Assay, Knockdown, Mutagenesis, Whisker Assay

Journal: Nature aging

Article Title: Sestrin is a key regulator of stem cell function and lifespan in response to dietary amino acids.

doi: 10.1038/s43587-020-00001-7

Figure Lengend Snippet: Fig. 5 | Sestrin overexpression in gut stem cells improves gut homeostasis. a,b, Ubiquitous overexpression of Sestrin reduced gut stem cell activity. a, Representative images from 30-day-old flies without (RU486−) or with (RU486+) induction. Actively dividing stem cells are indicated by arrows. Scale bar, 50 μm. b, Quantification of pH3-positive cells on days 10, 30 and 50. n = 20 guts (RU486−), n = 22 guts (RU486+), day 10; n = 23 guts (RU486−, RU486+), day 30; n = 24 guts (RU486−, RU486+), day 50. Generalized linear modelling analysis confirmed that Sestrin overexpression significantly reduced the increase in age-dependent stem cell activity (P = 0.012). c, Overexpression of Sestrin in enterocytes (5966GS) did not affect pH3-positive cell numbers. Day 30: n = 22 guts (RU486−), n = 21 guts (RU486+); day 46: n = 22 guts (RU486−), n = 20 guts (RU486+). d, In contrast, overexpression of Sestrin in gut stem cells (5961GS) significantly reduced stem cell activity. Day 30: n = 24 guts (RU486−), n = 25 guts (RU486+); day 46: n = 22 guts (RU486−), n = 23 guts (RU486+). e, Sestrin overexpression (OE) prolonged the G1 phase in gut stem cells. Fly-Fucci and Sestrin were driven by the esg-Gal4 driver. Sestrin overexpression significantly changed the cell cycle distribution of gut stem cells. n = 416 cells, wDah; n = 382 cells, Sesn OE. Chi-square test. f–h, Sestrin overexpression in gut stem cells decreased gut dysplasia (f,g) and the proportion of Smurf flies (h). f, Representative gut images from 50-day-old flies. DNA (DAPI) in blue. The gut epithelium is indicated by dashed lines. Scale bar, 20 μm. g, The difference in gut dysplasia was significant. n = 13 guts. h, Flies were 60 days old. n = 15 vials. Median, 25th and 75th percentiles, and Tukey whiskers are indicated in box-and-whisker plots (b–d,g,h). Outliers are shown as open circles. Statistics in b–d: two-tailed, Mann–Whitney test; statistics in g and h: unpaired, two-tailed t-test.

Article Snippet: Science 327, 1223-1228 3 n atu re research | rep o rtin g su m m ary O c to b e r 2 0 1 8 Anti pH3 (S10) rabbit polyclonal, Cell signaling, Cat# 9701.

Techniques: Over Expression, Activity Assay, Whisker Assay, Two Tailed Test, MANN-WHITNEY

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ph3 s10 rabbit polyclonal (Cell Signaling Technology Inc)

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