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gdf15 protein levels  (Boster Bio)


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    Boster Bio gdf15 protein levels
    Figure 1: Effect of oral intake of Plaquenil on <t>GDF15</t> and ghrelin plasma levels, and hunger scores in healthy volunteers. A) Time-dependent changes in plasma GDF15 levels after oral ingestion of Plaquenil or placebo in healthy volunteers (n ¼ 10; *P < 0.05: versus baseline [10 min], ##P < 0.01: versus the placebo condition; ANCOVA mixed model). Dashed line: trendline in the fasted state; dotted line: extrapolation from trendline in the fasted state. B and C) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and hungers scores measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values). D and E) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and ghrelin plasma levels measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values).All data were presented as mean SEM.
    Gdf15 Protein Levels, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity."

    Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

    Journal: Molecular metabolism

    doi: 10.1016/j.molmet.2024.102002

    Figure 1: Effect of oral intake of Plaquenil on GDF15 and ghrelin plasma levels, and hunger scores in healthy volunteers. A) Time-dependent changes in plasma GDF15 levels after oral ingestion of Plaquenil or placebo in healthy volunteers (n ¼ 10; *P < 0.05: versus baseline [10 min], ##P < 0.01: versus the placebo condition; ANCOVA mixed model). Dashed line: trendline in the fasted state; dotted line: extrapolation from trendline in the fasted state. B and C) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and hungers scores measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values). D and E) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and ghrelin plasma levels measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values).All data were presented as mean SEM.
    Figure Legend Snippet: Figure 1: Effect of oral intake of Plaquenil on GDF15 and ghrelin plasma levels, and hunger scores in healthy volunteers. A) Time-dependent changes in plasma GDF15 levels after oral ingestion of Plaquenil or placebo in healthy volunteers (n ¼ 10; *P < 0.05: versus baseline [10 min], ##P < 0.01: versus the placebo condition; ANCOVA mixed model). Dashed line: trendline in the fasted state; dotted line: extrapolation from trendline in the fasted state. B and C) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and hungers scores measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values). D and E) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and ghrelin plasma levels measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values).All data were presented as mean SEM.

    Techniques Used: Clinical Proteomics, Transformation Assay

    Figure 2: Distribution and characterization of GDF15 expressing epithelial cells in the proximal gut of normal-weight individuals and patients with obesity. A) Relative mRNA expression (efficiencyDDCt method) of GDF15 in resection specimens from the fundus (nNW ¼ 7, nOB ¼ 10), corpus (nNW ¼ 5, nOB ¼ 8), antrum (nNW ¼ 5, nOB ¼ 8), and jejunum (nNW ¼ 5, nOB ¼ 7) of normal-weight individuals and patients with obesity. (Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 in jejunal sections (10 mm) of a normal-weight individual and a patient with obesity. GDF15þ cells were stained with Cy3 (green) and nuclei were stained with DAPI (blue). Scale bars: 25 mm. C) Average number and intensity of GDF15þ cells in jejunal tissue sections from both normal-weight individuals and patients with obesity (nNW ¼ 5, nOB ¼ 5; two-tailed unpaired Student’s t-test). DeH) Representative double-immunofluorescence staining for GDF15 (green [Cy3 or Alexa594]) with markers for (D) goblet (MUC2: red [Cy5]), (E) Paneth (a-defensin 6: red [Cy5]) or (FeG) enteroendocrine (CHGA: red [Alexa488]) and ghrelin (red [Alexa488]) cells in jejunal sections (10 mm) from normal-weight individuals. Arrows indicate co-localization. Normal rabbit serum was used as negative control. Nuclei were labeled with DAPI (blue). Scale bars: 25 mm. NW: normal-weight, OB: obese. Data of figure A, C represent mean SEM and single values are plotted. *P < 0.05: versus jejunum in normal-weight individuals, $$$P < 0.001: versus jejunum in patients with obesity, #P < 0.05, ###P < 0.001: versus normal-weight individuals.
    Figure Legend Snippet: Figure 2: Distribution and characterization of GDF15 expressing epithelial cells in the proximal gut of normal-weight individuals and patients with obesity. A) Relative mRNA expression (efficiencyDDCt method) of GDF15 in resection specimens from the fundus (nNW ¼ 7, nOB ¼ 10), corpus (nNW ¼ 5, nOB ¼ 8), antrum (nNW ¼ 5, nOB ¼ 8), and jejunum (nNW ¼ 5, nOB ¼ 7) of normal-weight individuals and patients with obesity. (Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 in jejunal sections (10 mm) of a normal-weight individual and a patient with obesity. GDF15þ cells were stained with Cy3 (green) and nuclei were stained with DAPI (blue). Scale bars: 25 mm. C) Average number and intensity of GDF15þ cells in jejunal tissue sections from both normal-weight individuals and patients with obesity (nNW ¼ 5, nOB ¼ 5; two-tailed unpaired Student’s t-test). DeH) Representative double-immunofluorescence staining for GDF15 (green [Cy3 or Alexa594]) with markers for (D) goblet (MUC2: red [Cy5]), (E) Paneth (a-defensin 6: red [Cy5]) or (FeG) enteroendocrine (CHGA: red [Alexa488]) and ghrelin (red [Alexa488]) cells in jejunal sections (10 mm) from normal-weight individuals. Arrows indicate co-localization. Normal rabbit serum was used as negative control. Nuclei were labeled with DAPI (blue). Scale bars: 25 mm. NW: normal-weight, OB: obese. Data of figure A, C represent mean SEM and single values are plotted. *P < 0.05: versus jejunum in normal-weight individuals, $$$P < 0.001: versus jejunum in patients with obesity, #P < 0.05, ###P < 0.001: versus normal-weight individuals.

    Techniques Used: Expressing, Staining, Two Tailed Test, Negative Control, Labeling

    Figure 3: Effects of bitter generalists and intermediates on GDF15 or GLP-1 expression in jejunal crypts from patients with obesity. A) Effect of 4-hour stimulation with 100 mM HCQS on relative GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). B) Effect of 4-hour stimulation with 100 mM HCQS on relative GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). C) Effect of bitter generalists on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 6e9; Proc Mixed Model with Sidák correction for multiple comparisons). D) Effect of bitter intermediates on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e6; Proc Mixed Model with Sidák correction for multiple comparisons). E) Relative GDF15 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin. Azithromycin was removed after 4 h for the 24-hour timepoint (n ¼ 3e6; one-tailed paired Student’s t-test). F) Relative GLP-1 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin for 6 h (n ¼ 3; two-tailed paired Student’s t-test). All data are present as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus the vehicle group.
    Figure Legend Snippet: Figure 3: Effects of bitter generalists and intermediates on GDF15 or GLP-1 expression in jejunal crypts from patients with obesity. A) Effect of 4-hour stimulation with 100 mM HCQS on relative GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). B) Effect of 4-hour stimulation with 100 mM HCQS on relative GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). C) Effect of bitter generalists on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 6e9; Proc Mixed Model with Sidák correction for multiple comparisons). D) Effect of bitter intermediates on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e6; Proc Mixed Model with Sidák correction for multiple comparisons). E) Relative GDF15 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin. Azithromycin was removed after 4 h for the 24-hour timepoint (n ¼ 3e6; one-tailed paired Student’s t-test). F) Relative GLP-1 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin for 6 h (n ¼ 3; two-tailed paired Student’s t-test). All data are present as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus the vehicle group.

    Techniques Used: Expressing, Two Tailed Test, One-tailed Test

    Figure 4: Effects of bitter specialists on GDF15 mRNA or protein expression in primary jejunal crypts from patients with obesity. A) Effect of specialists on relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e7; Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 (green [Cy3]) in primary crypts treated with 1 mM gallic acid or vehicle after 24 h with 4 h stimulation. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. C) Average intensity of GDF15þ cells in primary crypts treated with 1 mM gallic acid or vehicle. Gallic acid was removed after 4 h of stimulation and staining was performed 20 h later (n ¼ 4; two-tailed paired Student’s t-test). D) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen for 6 h (n ¼ 3; one-tailed paired Student’s t-test). E) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen. Acetaminophen was removed after 4 h for the 24-hour timepoint (n ¼ 3; one-tailed paired Student’s t-test). F) Venn diagram of bitter compounds that affected GDF15 and/or GLP-1 mRNA expression after stimulation or primary crypts for 4 h. Asterisk (*) indicates an inhibitory effect on mRNA expression; the use of hashtags (#) signifies potential variations in the effects on GDF15 or GLP-1 mRNA, depending on the genotype of the patients. Data of figure A, C, D, E are present as mean SEM and single values are plotted. **P < 0.01, ***P < 0.001: versus the vehicle group.
    Figure Legend Snippet: Figure 4: Effects of bitter specialists on GDF15 mRNA or protein expression in primary jejunal crypts from patients with obesity. A) Effect of specialists on relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e7; Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 (green [Cy3]) in primary crypts treated with 1 mM gallic acid or vehicle after 24 h with 4 h stimulation. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. C) Average intensity of GDF15þ cells in primary crypts treated with 1 mM gallic acid or vehicle. Gallic acid was removed after 4 h of stimulation and staining was performed 20 h later (n ¼ 4; two-tailed paired Student’s t-test). D) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen for 6 h (n ¼ 3; one-tailed paired Student’s t-test). E) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen. Acetaminophen was removed after 4 h for the 24-hour timepoint (n ¼ 3; one-tailed paired Student’s t-test). F) Venn diagram of bitter compounds that affected GDF15 and/or GLP-1 mRNA expression after stimulation or primary crypts for 4 h. Asterisk (*) indicates an inhibitory effect on mRNA expression; the use of hashtags (#) signifies potential variations in the effects on GDF15 or GLP-1 mRNA, depending on the genotype of the patients. Data of figure A, C, D, E are present as mean SEM and single values are plotted. **P < 0.01, ***P < 0.001: versus the vehicle group.

    Techniques Used: Expressing, Staining, Labeling, Two Tailed Test, One-tailed Test

    Figure 5: Role of TAS2Rs and the motilin receptor in the effect of bitter agonists on GDF15 or GLP-1 expression in primary crypts from patients with obesity. A) The TAS2R antagonist, GIV3727 (110 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). B) GIV3727 (110 mM) did not block the effect of gallic acid (1 mM) on GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). C) GIV3727 (110 mM) did not block the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). D) The motilin receptor antagonist MA-2029 blocked the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). E) C12-O-AHL (0.15 mM) increased relative GDF15 mRNA expression in primary crypts from obese patients with TAS2R4(GG/CG) (n ¼ 9) but not TAS2R4(CC) (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). F) C12-O-AHL (0.15 mM) decreased GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R4(GG/CG) (n ¼ 9) but not the TAS2R4(CC) genotype (n ¼ 4). (Proc Mixed Model with Sidák correction for multiple comparisons). G) Aloin at 30 mM (white dots) and 100 mM (black dots) decreased relative GDF15 mRNA expression in primary crypts from obese patients with the TAS2R43(þ)GG/CG (n ¼ 9) but not the TAS2R43(þ)CC (n ¼ 6) or TAS2R43() (n ¼ 6) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). H) Aloin at 30 mM (white dots) and 100 mM (black dots) did not affect the relative GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R43(þ) GG/GC (n ¼ 9), TAS2R43(þ) CC (n ¼ 6), or TAS2R43() (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). IeK) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in jejunal sections (10 mm thickness) from normal-weight populations. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. MeO) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in primary jejunal crypt from patients with obesity. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. (L and P) Normal rabbit serum as a negative control. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. Data of figure AeH are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01, ###P < 0.001: bitter treatment antagonist, $P < 0.05: versus TAS2R4(CC) population, &P < 0.05: versus TAS2R43() population.
    Figure Legend Snippet: Figure 5: Role of TAS2Rs and the motilin receptor in the effect of bitter agonists on GDF15 or GLP-1 expression in primary crypts from patients with obesity. A) The TAS2R antagonist, GIV3727 (110 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). B) GIV3727 (110 mM) did not block the effect of gallic acid (1 mM) on GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). C) GIV3727 (110 mM) did not block the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). D) The motilin receptor antagonist MA-2029 blocked the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). E) C12-O-AHL (0.15 mM) increased relative GDF15 mRNA expression in primary crypts from obese patients with TAS2R4(GG/CG) (n ¼ 9) but not TAS2R4(CC) (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). F) C12-O-AHL (0.15 mM) decreased GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R4(GG/CG) (n ¼ 9) but not the TAS2R4(CC) genotype (n ¼ 4). (Proc Mixed Model with Sidák correction for multiple comparisons). G) Aloin at 30 mM (white dots) and 100 mM (black dots) decreased relative GDF15 mRNA expression in primary crypts from obese patients with the TAS2R43(þ)GG/CG (n ¼ 9) but not the TAS2R43(þ)CC (n ¼ 6) or TAS2R43() (n ¼ 6) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). H) Aloin at 30 mM (white dots) and 100 mM (black dots) did not affect the relative GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R43(þ) GG/GC (n ¼ 9), TAS2R43(þ) CC (n ¼ 6), or TAS2R43() (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). IeK) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in jejunal sections (10 mm thickness) from normal-weight populations. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. MeO) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in primary jejunal crypt from patients with obesity. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. (L and P) Normal rabbit serum as a negative control. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. Data of figure AeH are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01, ###P < 0.001: bitter treatment antagonist, $P < 0.05: versus TAS2R4(CC) population, &P < 0.05: versus TAS2R43() population.

    Techniques Used: Expressing, Blocking Assay, Labeling, Negative Control

    Figure 6: Role of the unfolded protein response pathway in the effect of bitter agonists on GDF15 and GLP1 mRNA expression in primary crypts from patients with obesity. A) Positive correlation between Log2 fold change of GDF15 and DDIT3 mRNA expression induced by bitter agonists (bitter: DB, AZI, GA, PHE, EM, ACE, BERB; n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). B) Positive correlation between Log2 fold change of GDF15 and ATF4 mRNA expression induced by bitter agonists from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). C) Positive correlation between Log2 fold change of DDIT3 and ATF4 mRNA expression induced by bitter agonist from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). D) ISRIB (5 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). E) ISRIB (5 mM) did not affect the effects of gallic acid (1 mM) on GLP-1 mRNA expression (n ¼ 2; Proc Mixed Model with Sidák correction for multiple comparisons). Data are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01: bitter treatment antagonist.
    Figure Legend Snippet: Figure 6: Role of the unfolded protein response pathway in the effect of bitter agonists on GDF15 and GLP1 mRNA expression in primary crypts from patients with obesity. A) Positive correlation between Log2 fold change of GDF15 and DDIT3 mRNA expression induced by bitter agonists (bitter: DB, AZI, GA, PHE, EM, ACE, BERB; n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). B) Positive correlation between Log2 fold change of GDF15 and ATF4 mRNA expression induced by bitter agonists from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). C) Positive correlation between Log2 fold change of DDIT3 and ATF4 mRNA expression induced by bitter agonist from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). D) ISRIB (5 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). E) ISRIB (5 mM) did not affect the effects of gallic acid (1 mM) on GLP-1 mRNA expression (n ¼ 2; Proc Mixed Model with Sidák correction for multiple comparisons). Data are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01: bitter treatment antagonist.

    Techniques Used: Expressing



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    Proteintech gdf15 levels
    The effects of ID8 implantation on muscle mass, fat mass, spleen mass, <t>GDF15</t> and gene expression of inflammation and atrogenes. Analysis of muscle mass at all time points in hindlimb muscles was completed (A, n = 22–24; soleus (SOL), extensor digitorum longus (EDL), plantaris (PLA), tibialis anterior (TA), gastrocnemius (GA) and quadriceps (QUAD)). Subcutaneous adipose mass in the inguinal fat depot (B, n = 9–12), serum GDF15 (C, n = 8–11) and spleen mass (D, n = 21–22) were also analyzed. mRNA content of inflammatory and atrophy markers interleukin-6 (IL-6), tumour necrosis factor – alpha (TNF-α), atrogin and muscle RING-finger protein-1 (MURF-1) were measured using quantitative PCR in the TA and diaphragm of all groups (E-H, n = 6–8). Results represent mean ± SD. Lettering denotes statistical significance when different from each other ( p < 0.05). C57BL/6J female mice ∼75 days post PBS injection as controls (CTRL); C57BL/6J female mice ∼45 days post ovarian cancer injection (45 Days); C57BL/6J female mice ∼75 days post ovarian cancer injection (75 Days); C57BL/6J female mice ∼90 days post ovarian cancer injection (90 Days). All data was analyzed using a one-way ANOVA or Kruskal–Wallis test when data did not fit normality. All ANOVAs were followed by a two-stage step-up method of Benjamini, Krieger and Yukutieli multiple comparisons test.
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    Figure 1: Effect of oral intake of Plaquenil on GDF15 and ghrelin plasma levels, and hunger scores in healthy volunteers. A) Time-dependent changes in plasma GDF15 levels after oral ingestion of Plaquenil or placebo in healthy volunteers (n ¼ 10; *P < 0.05: versus baseline [10 min], ##P < 0.01: versus the placebo condition; ANCOVA mixed model). Dashed line: trendline in the fasted state; dotted line: extrapolation from trendline in the fasted state. B and C) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and hungers scores measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values). D and E) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and ghrelin plasma levels measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values).All data were presented as mean SEM.

    Journal: Molecular metabolism

    Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

    doi: 10.1016/j.molmet.2024.102002

    Figure Lengend Snippet: Figure 1: Effect of oral intake of Plaquenil on GDF15 and ghrelin plasma levels, and hunger scores in healthy volunteers. A) Time-dependent changes in plasma GDF15 levels after oral ingestion of Plaquenil or placebo in healthy volunteers (n ¼ 10; *P < 0.05: versus baseline [10 min], ##P < 0.01: versus the placebo condition; ANCOVA mixed model). Dashed line: trendline in the fasted state; dotted line: extrapolation from trendline in the fasted state. B and C) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and hungers scores measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values). D and E) Oral intake of Plaquenil, but not placebo, resulted in a negative correlation between GDF15 plasma levels and ghrelin plasma levels measured between 0 and 90 min after administration. (n ¼ 10; Pearson correlation coefficient within individuals was transformed into Fisher z to calculate the average coefficient [r] and P values).All data were presented as mean SEM.

    Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

    Techniques: Clinical Proteomics, Transformation Assay

    Figure 2: Distribution and characterization of GDF15 expressing epithelial cells in the proximal gut of normal-weight individuals and patients with obesity. A) Relative mRNA expression (efficiencyDDCt method) of GDF15 in resection specimens from the fundus (nNW ¼ 7, nOB ¼ 10), corpus (nNW ¼ 5, nOB ¼ 8), antrum (nNW ¼ 5, nOB ¼ 8), and jejunum (nNW ¼ 5, nOB ¼ 7) of normal-weight individuals and patients with obesity. (Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 in jejunal sections (10 mm) of a normal-weight individual and a patient with obesity. GDF15þ cells were stained with Cy3 (green) and nuclei were stained with DAPI (blue). Scale bars: 25 mm. C) Average number and intensity of GDF15þ cells in jejunal tissue sections from both normal-weight individuals and patients with obesity (nNW ¼ 5, nOB ¼ 5; two-tailed unpaired Student’s t-test). DeH) Representative double-immunofluorescence staining for GDF15 (green [Cy3 or Alexa594]) with markers for (D) goblet (MUC2: red [Cy5]), (E) Paneth (a-defensin 6: red [Cy5]) or (FeG) enteroendocrine (CHGA: red [Alexa488]) and ghrelin (red [Alexa488]) cells in jejunal sections (10 mm) from normal-weight individuals. Arrows indicate co-localization. Normal rabbit serum was used as negative control. Nuclei were labeled with DAPI (blue). Scale bars: 25 mm. NW: normal-weight, OB: obese. Data of figure A, C represent mean SEM and single values are plotted. *P < 0.05: versus jejunum in normal-weight individuals, $$$P < 0.001: versus jejunum in patients with obesity, #P < 0.05, ###P < 0.001: versus normal-weight individuals.

    Journal: Molecular metabolism

    Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

    doi: 10.1016/j.molmet.2024.102002

    Figure Lengend Snippet: Figure 2: Distribution and characterization of GDF15 expressing epithelial cells in the proximal gut of normal-weight individuals and patients with obesity. A) Relative mRNA expression (efficiencyDDCt method) of GDF15 in resection specimens from the fundus (nNW ¼ 7, nOB ¼ 10), corpus (nNW ¼ 5, nOB ¼ 8), antrum (nNW ¼ 5, nOB ¼ 8), and jejunum (nNW ¼ 5, nOB ¼ 7) of normal-weight individuals and patients with obesity. (Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 in jejunal sections (10 mm) of a normal-weight individual and a patient with obesity. GDF15þ cells were stained with Cy3 (green) and nuclei were stained with DAPI (blue). Scale bars: 25 mm. C) Average number and intensity of GDF15þ cells in jejunal tissue sections from both normal-weight individuals and patients with obesity (nNW ¼ 5, nOB ¼ 5; two-tailed unpaired Student’s t-test). DeH) Representative double-immunofluorescence staining for GDF15 (green [Cy3 or Alexa594]) with markers for (D) goblet (MUC2: red [Cy5]), (E) Paneth (a-defensin 6: red [Cy5]) or (FeG) enteroendocrine (CHGA: red [Alexa488]) and ghrelin (red [Alexa488]) cells in jejunal sections (10 mm) from normal-weight individuals. Arrows indicate co-localization. Normal rabbit serum was used as negative control. Nuclei were labeled with DAPI (blue). Scale bars: 25 mm. NW: normal-weight, OB: obese. Data of figure A, C represent mean SEM and single values are plotted. *P < 0.05: versus jejunum in normal-weight individuals, $$$P < 0.001: versus jejunum in patients with obesity, #P < 0.05, ###P < 0.001: versus normal-weight individuals.

    Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

    Techniques: Expressing, Staining, Two Tailed Test, Negative Control, Labeling

    Figure 3: Effects of bitter generalists and intermediates on GDF15 or GLP-1 expression in jejunal crypts from patients with obesity. A) Effect of 4-hour stimulation with 100 mM HCQS on relative GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). B) Effect of 4-hour stimulation with 100 mM HCQS on relative GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). C) Effect of bitter generalists on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 6e9; Proc Mixed Model with Sidák correction for multiple comparisons). D) Effect of bitter intermediates on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e6; Proc Mixed Model with Sidák correction for multiple comparisons). E) Relative GDF15 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin. Azithromycin was removed after 4 h for the 24-hour timepoint (n ¼ 3e6; one-tailed paired Student’s t-test). F) Relative GLP-1 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin for 6 h (n ¼ 3; two-tailed paired Student’s t-test). All data are present as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus the vehicle group.

    Journal: Molecular metabolism

    Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

    doi: 10.1016/j.molmet.2024.102002

    Figure Lengend Snippet: Figure 3: Effects of bitter generalists and intermediates on GDF15 or GLP-1 expression in jejunal crypts from patients with obesity. A) Effect of 4-hour stimulation with 100 mM HCQS on relative GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). B) Effect of 4-hour stimulation with 100 mM HCQS on relative GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 5; two-tailed paired Student’s t-test). C) Effect of bitter generalists on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 6e9; Proc Mixed Model with Sidák correction for multiple comparisons). D) Effect of bitter intermediates on fold change in relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e6; Proc Mixed Model with Sidák correction for multiple comparisons). E) Relative GDF15 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin. Azithromycin was removed after 4 h for the 24-hour timepoint (n ¼ 3e6; one-tailed paired Student’s t-test). F) Relative GLP-1 secretion in the supernatant of primary crypts from patients with obesity stimulated with 1 mM azithromycin for 6 h (n ¼ 3; two-tailed paired Student’s t-test). All data are present as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus the vehicle group.

    Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

    Techniques: Expressing, Two Tailed Test, One-tailed Test

    Figure 4: Effects of bitter specialists on GDF15 mRNA or protein expression in primary jejunal crypts from patients with obesity. A) Effect of specialists on relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e7; Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 (green [Cy3]) in primary crypts treated with 1 mM gallic acid or vehicle after 24 h with 4 h stimulation. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. C) Average intensity of GDF15þ cells in primary crypts treated with 1 mM gallic acid or vehicle. Gallic acid was removed after 4 h of stimulation and staining was performed 20 h later (n ¼ 4; two-tailed paired Student’s t-test). D) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen for 6 h (n ¼ 3; one-tailed paired Student’s t-test). E) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen. Acetaminophen was removed after 4 h for the 24-hour timepoint (n ¼ 3; one-tailed paired Student’s t-test). F) Venn diagram of bitter compounds that affected GDF15 and/or GLP-1 mRNA expression after stimulation or primary crypts for 4 h. Asterisk (*) indicates an inhibitory effect on mRNA expression; the use of hashtags (#) signifies potential variations in the effects on GDF15 or GLP-1 mRNA, depending on the genotype of the patients. Data of figure A, C, D, E are present as mean SEM and single values are plotted. **P < 0.01, ***P < 0.001: versus the vehicle group.

    Journal: Molecular metabolism

    Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

    doi: 10.1016/j.molmet.2024.102002

    Figure Lengend Snippet: Figure 4: Effects of bitter specialists on GDF15 mRNA or protein expression in primary jejunal crypts from patients with obesity. A) Effect of specialists on relative GDF15 mRNA expression after 4 h of stimulation (n ¼ 4e7; Proc Mixed Model with Sidák correction for multiple comparisons). B) Representative single-immunofluorescence staining for GDF15 (green [Cy3]) in primary crypts treated with 1 mM gallic acid or vehicle after 24 h with 4 h stimulation. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. C) Average intensity of GDF15þ cells in primary crypts treated with 1 mM gallic acid or vehicle. Gallic acid was removed after 4 h of stimulation and staining was performed 20 h later (n ¼ 4; two-tailed paired Student’s t-test). D) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen for 6 h (n ¼ 3; one-tailed paired Student’s t-test). E) Relative GDF15 secretion in the supernatant of primary crypts stimulated with 3 mM acetaminophen. Acetaminophen was removed after 4 h for the 24-hour timepoint (n ¼ 3; one-tailed paired Student’s t-test). F) Venn diagram of bitter compounds that affected GDF15 and/or GLP-1 mRNA expression after stimulation or primary crypts for 4 h. Asterisk (*) indicates an inhibitory effect on mRNA expression; the use of hashtags (#) signifies potential variations in the effects on GDF15 or GLP-1 mRNA, depending on the genotype of the patients. Data of figure A, C, D, E are present as mean SEM and single values are plotted. **P < 0.01, ***P < 0.001: versus the vehicle group.

    Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

    Techniques: Expressing, Staining, Labeling, Two Tailed Test, One-tailed Test

    Figure 5: Role of TAS2Rs and the motilin receptor in the effect of bitter agonists on GDF15 or GLP-1 expression in primary crypts from patients with obesity. A) The TAS2R antagonist, GIV3727 (110 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). B) GIV3727 (110 mM) did not block the effect of gallic acid (1 mM) on GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). C) GIV3727 (110 mM) did not block the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). D) The motilin receptor antagonist MA-2029 blocked the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). E) C12-O-AHL (0.15 mM) increased relative GDF15 mRNA expression in primary crypts from obese patients with TAS2R4(GG/CG) (n ¼ 9) but not TAS2R4(CC) (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). F) C12-O-AHL (0.15 mM) decreased GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R4(GG/CG) (n ¼ 9) but not the TAS2R4(CC) genotype (n ¼ 4). (Proc Mixed Model with Sidák correction for multiple comparisons). G) Aloin at 30 mM (white dots) and 100 mM (black dots) decreased relative GDF15 mRNA expression in primary crypts from obese patients with the TAS2R43(þ)GG/CG (n ¼ 9) but not the TAS2R43(þ)CC (n ¼ 6) or TAS2R43() (n ¼ 6) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). H) Aloin at 30 mM (white dots) and 100 mM (black dots) did not affect the relative GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R43(þ) GG/GC (n ¼ 9), TAS2R43(þ) CC (n ¼ 6), or TAS2R43() (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). IeK) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in jejunal sections (10 mm thickness) from normal-weight populations. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. MeO) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in primary jejunal crypt from patients with obesity. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. (L and P) Normal rabbit serum as a negative control. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. Data of figure AeH are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01, ###P < 0.001: bitter treatment antagonist, $P < 0.05: versus TAS2R4(CC) population, &P < 0.05: versus TAS2R43() population.

    Journal: Molecular metabolism

    Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

    doi: 10.1016/j.molmet.2024.102002

    Figure Lengend Snippet: Figure 5: Role of TAS2Rs and the motilin receptor in the effect of bitter agonists on GDF15 or GLP-1 expression in primary crypts from patients with obesity. A) The TAS2R antagonist, GIV3727 (110 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). B) GIV3727 (110 mM) did not block the effect of gallic acid (1 mM) on GLP-1 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). C) GIV3727 (110 mM) did not block the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). D) The motilin receptor antagonist MA-2029 blocked the effect of azithromycin (75 mM) on GDF15 mRNA expression (efficiencyDDCt method) (n ¼ 4; Proc Mixed Model with Sidák correction for multiple comparisons). E) C12-O-AHL (0.15 mM) increased relative GDF15 mRNA expression in primary crypts from obese patients with TAS2R4(GG/CG) (n ¼ 9) but not TAS2R4(CC) (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). F) C12-O-AHL (0.15 mM) decreased GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R4(GG/CG) (n ¼ 9) but not the TAS2R4(CC) genotype (n ¼ 4). (Proc Mixed Model with Sidák correction for multiple comparisons). G) Aloin at 30 mM (white dots) and 100 mM (black dots) decreased relative GDF15 mRNA expression in primary crypts from obese patients with the TAS2R43(þ)GG/CG (n ¼ 9) but not the TAS2R43(þ)CC (n ¼ 6) or TAS2R43() (n ¼ 6) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). H) Aloin at 30 mM (white dots) and 100 mM (black dots) did not affect the relative GLP-1 mRNA expression in primary crypts from obese patients with the TAS2R43(þ) GG/GC (n ¼ 9), TAS2R43(þ) CC (n ¼ 6), or TAS2R43() (n ¼ 4) genotype (Proc Mixed Model with Sidák correction for multiple comparisons). IeK) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in jejunal sections (10 mm thickness) from normal-weight populations. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. MeO) Representative double-immunofluorescence images for GDF15 (green [Cy3]) and TAS2R4 (red [Cy5]), TAS2R43 (red [Cy5]) or TAS2R10 (red [Cy5]) in primary jejunal crypt from patients with obesity. Arrows indicate co-localization. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. (L and P) Normal rabbit serum as a negative control. Nuclei were labeled with DAPI (blue). Scale bars: 20 mm. Data of figure AeH are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01, ###P < 0.001: bitter treatment antagonist, $P < 0.05: versus TAS2R4(CC) population, &P < 0.05: versus TAS2R43() population.

    Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

    Techniques: Expressing, Blocking Assay, Labeling, Negative Control

    Figure 6: Role of the unfolded protein response pathway in the effect of bitter agonists on GDF15 and GLP1 mRNA expression in primary crypts from patients with obesity. A) Positive correlation between Log2 fold change of GDF15 and DDIT3 mRNA expression induced by bitter agonists (bitter: DB, AZI, GA, PHE, EM, ACE, BERB; n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). B) Positive correlation between Log2 fold change of GDF15 and ATF4 mRNA expression induced by bitter agonists from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). C) Positive correlation between Log2 fold change of DDIT3 and ATF4 mRNA expression induced by bitter agonist from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). D) ISRIB (5 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). E) ISRIB (5 mM) did not affect the effects of gallic acid (1 mM) on GLP-1 mRNA expression (n ¼ 2; Proc Mixed Model with Sidák correction for multiple comparisons). Data are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01: bitter treatment antagonist.

    Journal: Molecular metabolism

    Article Title: Bitter-tasting drugs tune GDF15 and GLP-1 expression via bitter taste or motilin receptors in the intestine of patients with obesity.

    doi: 10.1016/j.molmet.2024.102002

    Figure Lengend Snippet: Figure 6: Role of the unfolded protein response pathway in the effect of bitter agonists on GDF15 and GLP1 mRNA expression in primary crypts from patients with obesity. A) Positive correlation between Log2 fold change of GDF15 and DDIT3 mRNA expression induced by bitter agonists (bitter: DB, AZI, GA, PHE, EM, ACE, BERB; n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). B) Positive correlation between Log2 fold change of GDF15 and ATF4 mRNA expression induced by bitter agonists from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). C) Positive correlation between Log2 fold change of DDIT3 and ATF4 mRNA expression induced by bitter agonist from figure A (n ¼ 4e5/bitter treatment; Pearson correlation coefficient [r]; P value on graph). D) ISRIB (5 mM) blocked the effect of gallic acid (1 mM) on GDF15 mRNA expression (n ¼ 3; Proc Mixed Model with Sidák correction for multiple comparisons). E) ISRIB (5 mM) did not affect the effects of gallic acid (1 mM) on GLP-1 mRNA expression (n ¼ 2; Proc Mixed Model with Sidák correction for multiple comparisons). Data are presented as mean SEM and single values are plotted. *P < 0.05, **P < 0.01, ***P < 0.001: versus vehicle group, ##P < 0.01: bitter treatment antagonist.

    Article Snippet: GDF15 protein levels in the cell culture supernatant and cell lysate were measured using a Human GDF-15 ELISA Kit (Boster Biological Technology), following the manufacturer’s protocol.

    Techniques: Expressing

    The effects of ID8 implantation on muscle mass, fat mass, spleen mass, GDF15 and gene expression of inflammation and atrogenes. Analysis of muscle mass at all time points in hindlimb muscles was completed (A, n = 22–24; soleus (SOL), extensor digitorum longus (EDL), plantaris (PLA), tibialis anterior (TA), gastrocnemius (GA) and quadriceps (QUAD)). Subcutaneous adipose mass in the inguinal fat depot (B, n = 9–12), serum GDF15 (C, n = 8–11) and spleen mass (D, n = 21–22) were also analyzed. mRNA content of inflammatory and atrophy markers interleukin-6 (IL-6), tumour necrosis factor – alpha (TNF-α), atrogin and muscle RING-finger protein-1 (MURF-1) were measured using quantitative PCR in the TA and diaphragm of all groups (E-H, n = 6–8). Results represent mean ± SD. Lettering denotes statistical significance when different from each other ( p < 0.05). C57BL/6J female mice ∼75 days post PBS injection as controls (CTRL); C57BL/6J female mice ∼45 days post ovarian cancer injection (45 Days); C57BL/6J female mice ∼75 days post ovarian cancer injection (75 Days); C57BL/6J female mice ∼90 days post ovarian cancer injection (90 Days). All data was analyzed using a one-way ANOVA or Kruskal–Wallis test when data did not fit normality. All ANOVAs were followed by a two-stage step-up method of Benjamini, Krieger and Yukutieli multiple comparisons test.

    Journal: Molecular Metabolism

    Article Title: Muscle weakness and mitochondrial stress occur before severe metastasis in a novel mouse model of ovarian cancer cachexia

    doi: 10.1016/j.molmet.2024.101976

    Figure Lengend Snippet: The effects of ID8 implantation on muscle mass, fat mass, spleen mass, GDF15 and gene expression of inflammation and atrogenes. Analysis of muscle mass at all time points in hindlimb muscles was completed (A, n = 22–24; soleus (SOL), extensor digitorum longus (EDL), plantaris (PLA), tibialis anterior (TA), gastrocnemius (GA) and quadriceps (QUAD)). Subcutaneous adipose mass in the inguinal fat depot (B, n = 9–12), serum GDF15 (C, n = 8–11) and spleen mass (D, n = 21–22) were also analyzed. mRNA content of inflammatory and atrophy markers interleukin-6 (IL-6), tumour necrosis factor – alpha (TNF-α), atrogin and muscle RING-finger protein-1 (MURF-1) were measured using quantitative PCR in the TA and diaphragm of all groups (E-H, n = 6–8). Results represent mean ± SD. Lettering denotes statistical significance when different from each other ( p < 0.05). C57BL/6J female mice ∼75 days post PBS injection as controls (CTRL); C57BL/6J female mice ∼45 days post ovarian cancer injection (45 Days); C57BL/6J female mice ∼75 days post ovarian cancer injection (75 Days); C57BL/6J female mice ∼90 days post ovarian cancer injection (90 Days). All data was analyzed using a one-way ANOVA or Kruskal–Wallis test when data did not fit normality. All ANOVAs were followed by a two-stage step-up method of Benjamini, Krieger and Yukutieli multiple comparisons test.

    Article Snippet: GDF15 (Growth differentiation factor 15) levels were analyzed in serum using the mouse GDF-15 DuoSet ELISA kit according to the manufacturer's instructions (R&D Systems DY6385).

    Techniques: Expressing, Muscles, Real-time Polymerase Chain Reaction, Injection