Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Hydrangea serrata extract attenuates PM-exacerbated airway inflammation in the CARAS model by modulating the IL-33/ST2/NF-κB signaling pathway.

doi: 10.1016/j.biopha.2024.116596

Figure Lengend Snippet: Fig. 6. HSE regulated IL-4, IL-5, and histamine in PM-exacerbated CARAS mice. (A,B) Levels of IL-4 and IL-5 in NLF. (C,D) Levels of IL-4 and IL-5 in BALF. (E) Levels of histamine in BALF. (G) Levels of IL-33 in BALF. All levels were determined using ELISA kits. Values are presented as mean ± SEM. Significant differences at ###p<0.001 and ##p<0.01 compared with the control group; ***p<0.001, **p<0.01, and *p<0.05 compared with the CARAS/PM group. CARAS, combined allergic rhinitis and asthma syndrome; Dexa, dexamethasone; HSE, Hydrangea serrata extract; OVA, ovalbumin; PM, particulate matter.

Article Snippet: Thus, IL-33 in BALF was measured using an ELISA kit (M33000, R&D Systems Inc., Minneapolis, MN, USA) according to the manufacturer’s guidelines.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: JCI Insight

Article Title: Elevation of master autophagy regulator Tfeb in osteoblast lineage cells increases bone mass and strength

doi: 10.1172/jci.insight.191688

Figure Lengend Snippet: ( A – E ) Skeletal phenotype analysis of 4.5-month-old male Tfeb CRa mice and their littermate Cre controls (Osx1-Cre only [red dots], Osx1-Cre CRa [blue dots], Osx1-Cre sgRNA Tfeb [gray dots]). ( A – C ) Dynamic histomorphometry was performed on femurs. n = 5–6 mice per group. ( A ) Quantification of mineralizing surface per bone surface (MS/BS), mineral apposition rate (MAR), and bone formation rate per bone surface (BFR/BS) at the periosteal surface. ( B ) Representative histological cross section showing calcein labeling at the femoral diaphysis (GFP filter). Scale bars represent 400 μm. ( C ) Trabecular MAR (Tb. MAR). ( D and E ) Sp7 and Col1a1 ( D ) and Acp5 and Ctsk ( E ) mRNA levels were measured in L5 by qRT-PCR and normalized to mouse Actb . n = 18 mice per group (please see Methods). ( F and G ) Bone marrow cells were isolated from femurs and tibias of 5-month-old female mice and differentiated into osteoblasts using osteogenic media. ( F ) Alizarin red stain of mineral apposition. ( G ) BrdU analysis of proliferating cells. n = 4 wells per group. ( H ) Circulating sclerostin levels of 5- and 12-month-old male (square) and female (circle) Tfeb CRa and control mice were measured using ELISA. n = 5–7 mice per group. ( I ) Sclerostin and actin levels were measured in cortical bone (humeri shafts) of Tfeb CRa and control mice via Western blot. Immunoblot of 5 mice per group is shown. Quantification was done with n = 10–11 mice per group. The bar graph indicates sclerostin levels normalized to actin. Bars indicate mean ± SD. Indicated P values were calculated by unpaired t test for equal or unequal variance ( A -Ps.MAR).

Article Snippet: Circulating sclerostin levels were measured using Mouse/Rat SOST/Sclerostin Quantikine ELISA Kit (R&D Systems, MSST00).

Techniques: Labeling, Quantitative RT-PCR, Isolation, Staining, Control, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: JCI Insight

Article Title: Elevation of master autophagy regulator Tfeb in osteoblast lineage cells increases bone mass and strength

doi: 10.1172/jci.insight.191688

Figure Lengend Snippet: ( A – C ) Serial spine ( A ), femur ( B ), and global (whole body) ( C ) DXA BMD measurements were collected from female Tfeb CRa mice ( n = 6) and their littermate Cre controls (Osx1-Cre only, Osx1-Cre sgRNA Tfeb mice, n = 8) between 3 and 12 months of age. Error bars indicate SD. * P < 0.05 as calculated by unpaired t test at each time point. Indicated P values were calculated by repeated measures model (detailed in the Methods section) comparing the difference in genotypes at 3 versus 9 months or 9 versus 12 months. ( D and E ) μCT analysis of Ct.Th ( D ) and trabecular BV/TV ( E ) performed on lumbar vertebrae 4 (spine) and femurs from 12-month-old female Tfeb CRa mice ( n = 6) and littermate controls (Osx1-Cre only [red dots], Osx1-Cre sgRNA Tfeb [gray dots], n = 8). ( F ) ELISA measurements of P1NP and TRAcP 5b ( Tfeb CRa n = 6, controls n = 7). ( G – J ) Histomorphometric analysis of vertebral trabecular bone of Tfeb CRa mice ( n = 5) and their Cre controls ( n = 8). ( G ) MS/BS, BFR/BS. ( H ) Osteoblast surface per bone perimeter (Ob.S/B.Pm), osteoblast number per bone perimeter (N.Ob/B.Pm). ( I ) Osteoclast surface per bone perimeter (Oc.S/B.Pm), osteoclast number per bone perimeter (N.Oc/B.Pm). ( J ) TRAcP 5b– and toluidine blue–stained or unstained (labeled with calcein and alizarin red) sections imaged with 40× original magnification. Scale bars represent 50 μm. ( K and L ) Bglap and Sp7 ( K ) and Acp5 and Ctsk ( L ) mRNA levels measured in lumbar vertebrae 5 (spine) of Tfeb CRa mice ( n = 7) and controls ( n = 6) using qRT-PCR and normalized to mouse Actb . Bars indicate mean ± SD. Indicated P values were calculated by unpaired t test for equal or unequal variance ( E -femur and spine, K - Sp7 ) or rank sum test (if data are not normally distributed, D -femur, H –N.Ob/B.Pm).

Article Snippet: Circulating sclerostin levels were measured using Mouse/Rat SOST/Sclerostin Quantikine ELISA Kit (R&D Systems, MSST00).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Labeling, Quantitative RT-PCR

Journal: Food & Nutrition Research

Article Title: Cudrania tricuspidata water extract improved obesity-induced hepatic insulin resistance in db/db mice by suppressing ER stress and inflammation

doi: 10.3402/fnr.v59.29165

Figure Lengend Snippet: Levels of glucose, insulin, glucagon, and leptin in the C57BL/6J- db/db mice with dietary supplementation of Cudrania tricuspidata water extract

Article Snippet: Serum insulin, glucagon, and leptin were respectively measured with an Ultra Sensitive Mouse Insulin ELISA kit (Crystal Chem, Downers Grove, IL, USA), Glucagon Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA), and Mouse/Rat Leptin Quantikine ELISA kit (R&D Systems) according to the manufacturer's protocols.

Techniques: Control

Journal: bioRxiv

Article Title: PPARG in osteocytes is essential for sclerostin expression, bone mass, marrow adiposity and TZD-induced bone loss

doi: 10.1101/2020.09.13.295378

Figure Lengend Snippet: Correlation between PPARG and sclerostin levels in osteocytes. A. and B. Levels of PPARG KO and sclerostin proteins in OT isolated from 6.5 mo old male γOT KO and Ctrl mice and either analyzed KO on separate (A) or the same (B) Western blot membrane. (γOT KO n=5; Ctrl n=4). C) Pearson correlation analysis of PPARG and sclerostin protein levels shown in panel B. Coefficient of determination (R 2 =0.9821) was calculated based on bands density assessed by Image J and normalized to b-actin. D. KO Sclerostin levels measured by ELISA in sera of γOT KO (n=4) and Ctrl (n=7) 6 mo old male mice. Statistical significance was calculated using parametric unpaired Student’s t test.

Article Snippet: Sclerostin levels after depletion were quantified using SOST ELISA kit (Quantikine ELISA, R&D Systems, Cat#MSST00).

Techniques: Isolation, Western Blot, Membrane, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: PPARG in osteocytes is essential for sclerostin expression, bone mass, marrow adiposity and TZD-induced bone loss

doi: 10.1101/2020.09.13.295378

Figure Lengend Snippet: PPARG positively regulates sclerostin expression. A. Schematic positioning of JASPAR projected PPARG response elements (PPREs) upstream of sclerostin transcription start site (TSS). B. Chromatin immunoprecipitation (ChIP) assay end point agarose gel image (full image of the gel is presented in Suppl. Fig. 1). ChIP was performed on osteocyte-like MLO-Y4 cells targeting PPRE3 and PPRE14/15. Cells were treated for 24h with either vehicle (V), or 1µM rosiglitazone (R) or combination of 1µM R and 10µM GW9662 antagonist (R+GW) and ChIP assays were performed, as described in Material and Methods. S – sonicated lysate (no antibody pulldown). C. Expression of Sost mRNA in MLO-Y4 cells treated for 3 days with either vehicle (V), or rosiglitazone (R), or combination of rosiglitazone and GW9662 (R+GW) at the same doses as in (B). One-way ANOVA followed by Tukey’s post hoc analysis was performed for significance calculation. D. Western blot of sclerostin protein levels in MLO-Y4 cells treated as in (C). Sclerostin protein levels were normalized to β-actin levels measured as band density using Image J and relative levels of expression had been calculated and shown below Western blot images.

Article Snippet: Sclerostin levels after depletion were quantified using SOST ELISA kit (Quantikine ELISA, R&D Systems, Cat#MSST00).

Techniques: Expressing, Chromatin Immunoprecipitation, Agarose Gel Electrophoresis, Sonication, Western Blot

Journal: bioRxiv

Article Title: PPARG in osteocytes is essential for sclerostin expression, bone mass, marrow adiposity and TZD-induced bone loss

doi: 10.1101/2020.09.13.295378

Figure Lengend Snippet: Osteocyte derived sclerostin positively contributes to marrow adipocyte differentiation. A. Schematic showing experimental design of co-culture of BMSC with intact or sclerostin depleted conditioned medium (CM). One group of adherent BMSC culture received IgG depleted CM from primary osteocytes (control group) while the other group received sclerostin depleted CM (group of interest). B. ELISA measurements of sclerostin level in CM after anti-SOST antibody mediated depletion. C. Expression of adipocytic and osteoblastic gene markers in adherent BMSCs treated with CM from primary osteocytes IgG or sclerostin (Ab-Scl) depleted. Statistical significance was calculated using parametric unpaired Student’s t test.

Article Snippet: Sclerostin levels after depletion were quantified using SOST ELISA kit (Quantikine ELISA, R&D Systems, Cat#MSST00).

Techniques: Derivative Assay, Co-Culture Assay, Control, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Nature Communications

Article Title: Cancer cachexia in STK11/LKB1 -mutated non-small cell lung cancer is dependent on tumor-secreted GDF15

doi: 10.1038/s41467-026-68702-y

Figure Lengend Snippet: A List of non-cachexia NSCLC lines with wild-type STK11/LKB1 and cachexia NSCLC lines with mutant STK11/LKB1 from which tumors and serum have been obtained from previous work when lines were xenotransplanted in vivo . B – H Tumors were harvested, pooled and subjected to quantitative RT-PCR for human GDF15 ( B , E ) and mouse ( C , H ) Gdf15 mRNA normalized to β-actin. Serum was subjected to the human ( D – F ) or mouse ( G , H ) GDF15 ELISA as described in Methods. Data are shown as mean ± SEM ( B – D , G ) or as a scatter plot ( E , F , H ) or relative to the average non-cachexia cohort ( B , C , E , H ) or the actual measurements ( D , F , G ). P was calculated based on unpaired 2-tailed t test ( B and C ) or 1-way ANOVA followed by Dunnett’s multiple-comparison test ( D and G ), or from linear regression of GDF15 mRNA levels with serum GDF15 level ( E , F , H ). n.s. not significant. 6 replicates for each point except n = 4 for H411, n = 3 for HCC15, n = 3 for H1944, n = 5 for H1573. Source data are provided as a Source Data file.

Article Snippet: Mouse GDF15 in serum was measured using a mouse/rat GDF15 Quantikine ELISA kit (R&D Systems, #MGD150) following the manufacturer’s instructions.

Techniques: Mutagenesis, In Vivo, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Nature Communications

Article Title: Cancer cachexia in STK11/LKB1 -mutated non-small cell lung cancer is dependent on tumor-secreted GDF15

doi: 10.1038/s41467-026-68702-y

Figure Lengend Snippet: A – M Chow-fed 13-week-old NOD/SCID male mice ( n = 6 per group) were injected s.c. with 200 μl PBS in the absence or presence of 1 ×10 7 cells from parental H1573, H1573 ΔGFP , or H1573 ΔGDF15 lines as described in Methods. Tumor cells before injection (cells) and tumors at sacrifice (tumor) were processed for immunoblot analysis with the indicated antibody, and the samples derive from the same experiment but different gels for GDF15 and Actin ( A ). Longitudinal or endpoint measurements of tumor volume ( B ), tumor weight ( C ), food intake ( G ), body weight ( H ), tumor-free body weight ( I ), fat mass ( J ), lean mass ( K ), tumor-free lean mass ( L ), and forelimb grip strength ( M ) were obtained as described in Methods. Serum at sacrifice was subjected to the human GDF15 ( D ), mouse GDF15 ( E ) or mouse leptin ( F ) ELISA as described in Methods. Data are shown as mean ± SEM of the actual measurements ( B , C – G , M ) or relative to their day 0 values ( H – L ). P was calculated using 1-way ( C – G , I , L , M ) or 2-way ( B , H , J , K ) ANOVA followed by Dunnett’s multiple-comparison test for significant differences from the H1573 ΔGDF15 cohort. n.s. not significant; M = GDF15 mature protein; P = GDF15 proprotein. Source data are provided as a Source Data file.

Article Snippet: Mouse GDF15 in serum was measured using a mouse/rat GDF15 Quantikine ELISA kit (R&D Systems, #MGD150) following the manufacturer’s instructions.

Techniques: Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Nature Communications

Article Title: Cancer cachexia in STK11/LKB1 -mutated non-small cell lung cancer is dependent on tumor-secreted GDF15

doi: 10.1038/s41467-026-68702-y

Figure Lengend Snippet: A – J NOD/SCID mice were treated as described in Fig. ( n = 6 per group). Mice were sacrificed with epididymal white adipose tissue (eWAT) ( A ), inguinal white adipose tissue (iWAT) ( B ), gastrocnemius muscle ( F ), and quadriceps muscle ( G ) weighed. eWAT and gastrocnemius muscle were harvested and processed for H&E ( C ) or Laminin ( H ) staining, respectively (scale bar 100 μm). Subsequently, measurements of adipocyte area ( D , E ) ( n = 4) and myocyte ( H , I ) ( n = 3 for PBS, n = 4 for H1573, n = 5 for H1573 ΔGFP and H1573 ΔGDF15 ) cross-sectional area were obtained. Gastrocnemius muscle was harvested and subjected to quantitative RT-PCR analysis of mouse mRNA levels of the indicated genes ( n = 6 for non-tumor bearing mice group, n = 5 for tumor bearing mice group) ( J ). Data are shown as mean ± SEM of the actual measurements ( A , B , D , F , G , and I ), or as scattered plots with regression line and 95% confidence interval of relative values ( J ). P was calculated using 1-way ( A , B , D – G , I ) followed by Dunnett’s multiple-comparison test for significant differences from the H1573 ΔGDF15 cohort. P was calculated using Pearson correlation coefficients, two-tailed ( J ). n.s. not significant. Source data are provided as a Source Data file.

Article Snippet: Mouse GDF15 in serum was measured using a mouse/rat GDF15 Quantikine ELISA kit (R&D Systems, #MGD150) following the manufacturer’s instructions.

Techniques: Staining, Quantitative RT-PCR, Comparison, Two Tailed Test

Journal: Nature Communications

Article Title: Cancer cachexia in STK11/LKB1 -mutated non-small cell lung cancer is dependent on tumor-secreted GDF15

doi: 10.1038/s41467-026-68702-y

Figure Lengend Snippet: A – N Chow-fed 11–12-week-old NOD/SCID male mice ( n = 9 per group) were injected to left lung with 30 µl of 25% growth factor reduced Matrigel in dye-free RPMI medium in the absence or presence of 1 ×10 6 cells from H1573 ΔGFP , or H1573 ΔGDF15 lines as described in Methods. Longitudinal or endpoint measurements of tumor area ( A , B ), food intake ( E ), body weight ( F ), fat mass ( G ), lean mass ( H ), or forelimb grip strength ( I ) were obtained as described in Methods. Serum at sacrifice was subjected to the human ( C ) or mouse ( D ) GDF15 ELISA as described in Methods. Indicated muscles ( J ) or fat pads ( M ) were weighed. eWAT and gastrocnemius muscle were harvested and processed for H&E or Laminin staining, respectively ( L ) (scale bar 100 μm). Subsequently, measurements of myocyte ( K ) and adipocyte ( N ) cross-sectional area were obtained ( n = 6 for non-tumor bearing mice group, n = 8 for H1573 ΔGFP , n = 7 for H1573 ΔGDF15 ). Data are shown as mean ± SEM of the actual measurements ( B , C – E , I – K , M , N ) or relative to their day 0 values ( F – H ). P was calculated using 1-way ( B – E , I – K , M , N ) or 2-way ( F – H ) ANOVA followed by Dunnett’s multiple-comparison test for significant differences from the H1573 ΔGDF15 cohort. n.s. not significant. Source data are provided as a Source Data file.

Article Snippet: Mouse GDF15 in serum was measured using a mouse/rat GDF15 Quantikine ELISA kit (R&D Systems, #MGD150) following the manufacturer’s instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Muscles, Staining, Comparison

Journal: Nature Communications

Article Title: Cancer cachexia in STK11/LKB1 -mutated non-small cell lung cancer is dependent on tumor-secreted GDF15

doi: 10.1038/s41467-026-68702-y

Figure Lengend Snippet: A – P Chow-fed 12-13-week-old NOD/SCID male mice ( n = 6) were injected s.c. with 200 μl PBS containing 1 ×10 7 cells from parental H1573 human NSCLC line on day 0. A – G Starting on day 7 or day 15 ( H – P ), mice were injected with s.c. 150 μl PBS in the absence or presence of 10 mg/kg IgG or GDF15 antibody weekly. Longitudinal or endpoint measurements of tumor volume ( B , I ), tumor weight ( C , J ), tumor-free body weight ( E , L ), fat mass ( G , M ), tumor-free lean mass ( F , O ), eWAT weight ( N ), and Gastrocnemius weight ( P ) were obtained as described in Methods. Serum at sacrifice was processed for human GDF15 ELISA ( D , K ) as described in Methods. Data are shown as mean ± SEM of the actual measurements ( B – D , I – K , N , P ) or relative to their day 0 values ( E – G , L – M , O ). P was calculated using 1-way ( C – F , J – L , N – P ) or 2-way ( B , G , I and M ) ANOVA followed by Dunnett’s multiple-comparison test for significant differences from the GDF15 antibody cohort. n.s. not significant. Source data are provided as a Source Data file.

Article Snippet: Mouse GDF15 in serum was measured using a mouse/rat GDF15 Quantikine ELISA kit (R&D Systems, #MGD150) following the manufacturer’s instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Nature Communications

Article Title: Cancer cachexia in STK11/LKB1 -mutated non-small cell lung cancer is dependent on tumor-secreted GDF15

doi: 10.1038/s41467-026-68702-y

Figure Lengend Snippet: A – N Chow-fed 13-week-old NOD/SCID male mice ( n = 6 per group) were injected s.c. with 200 μl PBS in the absence or presence of 1 × 10 7 cells from parental H2122, H2122 ΔGFP , or H2122 ΔGDF15 lines as described in Methods. Tumor cells before injection (cells) and tumors at sacrifice (tumors) were processed for immunoblot analysis with the indicated antibody, and the samples derive from the same experiment but different gels for GDF15 and Actin ( A ). Longitudinal or endpoint measurements of tumor volume ( B ), tumor weight ( C ), food intake ( F ), tumor-free body weight ( G ), tumor-free lean mass ( H ), or fat mass ( K ) were obtained as described in Methods. At sacrifice, serum was subjected to the human ( D ) or mouse ( E ) GDF15 ELISA and gastrocnemius muscle and epididymal white adipose tissue were harvested and processed for Laminin ( I ) or H&E ( L ) staining, respectively (scale bar 100 μm). Subsequently, measurements of myocyte ( J ) ( n = 4 for non-tumor bearing mice group, n = 4 parental for H2122 and H2122 ΔGFP , n = 5 for H2122 ΔGDF15 ) and adipocyte ( M , N ) cross-sectional area from 3 mice per cohort were obtained as described in Methods. Data are shown as mean ± SEM of the actual measurements ( B – F , J , M , N ) or relative to their day 0 values ( G , H , K ). P was calculated using 1-way ( C – H , J , M , N ) or 2-way ( B , K ) ANOVA followed by Dunnett’s multiple-comparison test for significant differences from the H2122 ΔGDF15 cohort. n.s. not significant; M = GDF15 mature protein; P = GDF15 proprotein. Source data are provided as a Source Data file.

Article Snippet: Mouse GDF15 in serum was measured using a mouse/rat GDF15 Quantikine ELISA kit (R&D Systems, #MGD150) following the manufacturer’s instructions.

Techniques: Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Comparison

Journal: Nature Communications

Article Title: Cancer cachexia in STK11/LKB1 -mutated non-small cell lung cancer is dependent on tumor-secreted GDF15

doi: 10.1038/s41467-026-68702-y

Figure Lengend Snippet: A – J Chow-fed 12-week-old C57BL6J male mice ( n = 6) were injected s.c. with 200 μl PBS containing in the absence or presence of 1 ×10 6 cells from the mouse NSCLC KP or KPL lines on day 0 followed by weekly injections of 150 μl PBS in the absence or presence of 10 mg/kg IgG or GDF15 antibody. Longitudinal or endpoint measurements of tumor volume ( A ), tumor weight ( B ), food intake ( C ), tumor-free body weight ( D ), fat mass ( E ), and tumor-free lean mass ( H ) were obtained as described in Methods. At sacrifice, eWAT and gastrocnemius were harvested and processed for H&E ( F ) or Laminin ( I ) staining, respectively (scale bar 100 μm). Subsequently, measurements of adipocyte cross-sectional area ( G ) ( n = 4 mice) and myocyte ( J ) cross-sectional area ( n = 4 for KP, n = 3 for KPL tumor bearing mice without treatment, and n = 4 for KPL tumor bearing mice treated with IgG or GDF15 antibody) were obtained. Data are shown as mean ± SEM of the actual measurements ( A – D , G and J ) or relative to their day 0 values ( D – E , H ). P was calculated using 1-way ( B – D , G , H , J ) or 2-way ( A and E ) ANOVA followed by Dunnett’s multiple-comparison test for significant differences from the KPL + GDF15 antibody cohorts. n.s. not significant. Source data are provided as a Source Data file.

Article Snippet: Mouse GDF15 in serum was measured using a mouse/rat GDF15 Quantikine ELISA kit (R&D Systems, #MGD150) following the manufacturer’s instructions.

Techniques: Injection, Staining, Comparison

Journal: Nature Communications

Article Title: Cancer cachexia in STK11/LKB1 -mutated non-small cell lung cancer is dependent on tumor-secreted GDF15

doi: 10.1038/s41467-026-68702-y

Figure Lengend Snippet: A , B Expression and secretion of GDF15 from patient-derived NSCLC cell line reconstituted with wild-type STK11/LKB1 . Parental H1437, H1437 CTL , or H1437 STK11 cells were harvested ( A , J ) or incubated with medium D in the presence of 0.01–10 mM glucose followed by harvesting ( B ). D - I Effect of STK11/LKB1 reconstitution on cachexia phenotype. Chow-fed 15-week-old NOD/SCID male mice ( n = 6 per group) were injected s.c. with 200 μl PBS in the absence or presence of 1 × 10 7 cells from parental H1437, H1437 CTL , H1437 STK11 , or another H1437 STK11 monoclonal line as described in Methods. Longitudinal or endpoint measurements of tumor volume ( D ), food intake ( F ), tumor-free body weight ( G ), fat mass ( H ), or tumor-free lean mass ( I ) were obtained as described in Methods. Tumors at sacrifice were processed for immunoblot analysis with the indicated antibody as described in Methods ( C ). At sacrifice, serum was subjected to the human GDF15 ELISA as described in Methods ( E ). H1437 CTL , or H1437 STK11 cells stably expressed control vector or human GDF15-3xFlag protein, or parental H1437, H1437 ∆GFP or H1437 ∆ATF4 were harvested ( K , M ) or incubated with medium D in the presence of 10 mM or 0.1 mM glucose followed by harvesting ( L ). Medium and cell lysate were subjected to immunoblot analysis with the indicated antibody or quantitative RT-PCR with indicated primer, respectively as described in Methods. Data are shown as mean ± SEM of the actual measurements ( D – F ) or relative to day 0 values ( G – I ) or values relative to control group ( J , M ). Samples for quantitative RT-PCR analysis of gene expression were from 3 biological replicates per group ( J , M ). P was calculated using 1-way ( E – G , I – J , M ) or 2-way ( D , H ) ANOVA followed by Dunnett’s multiple-comparison test for significant differences from the H1437 parental cohort. n.s. not significant; M = GDF15 mature protein; P = GDF15 proprotein. The samples derive from the same experiment but different gels for different antibodies ( A – C , K , L ). Experiment was repeated at least 3 times with similar results ( A – C ). Source data are provided as a Source Data file.

Article Snippet: Mouse GDF15 in serum was measured using a mouse/rat GDF15 Quantikine ELISA kit (R&D Systems, #MGD150) following the manufacturer’s instructions.

Techniques: Expressing, Derivative Assay, Incubation, Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Stable Transfection, Control, Plasmid Preparation, Quantitative RT-PCR, Gene Expression, Comparison

Journal: European Burn Journal

Article Title: The Role of Angiopoietin-2 in Post-Burn Pneumonia

doi: 10.3390/ebj7010001

Figure Lengend Snippet: Early elevation of Angiopoietin-2 and Ang-2/1 ratio in patient serum correlates with 30-day mortality. Burn patients were prospectively enrolled and serum was isolated and evaluated via ELISA for Angiopoietin-1 and 2. Dark blue dots represent patients who did not develop pneumonia within 30 days of burn, and red dots represent patients who did develop pneumonia within 30 days of burn. ( A ) Post-burn day 2 (PBD2) serum Ang-2 and ( B ) PBD2 Ang-2/1 ratio for patients that did not (−) and did (+) develop pneumonia within 30 days of injury (( A ) p < 0.05, ( B ) p < 0.01). ( C ) Post-burn day 3 (PBD3) serum Ang-2 and ( D ) PBD3 Ang-2/1 ratio (( C ) p < 0.0001, ( D ) p < 0.01). Significance was analyzed via Mann–Whitney U-test, ** p < 0.01, **** p < 0.0001, PBD2 (−) n = 21, (+) n = 10, PBD3 (−) n = 23, (+) n = 7.

Article Snippet: Plasma Angiopoietin-2 levels were determined using Mouse & Rat Angiopoietin-2 Quantikine ELISA Kit (R&D Systems Cat. MANG20) using a 1:40 dilution of plasma in technical duplicate and reported as ng of Ang-2/mL of plasma.

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: European Burn Journal

Article Title: The Role of Angiopoietin-2 in Post-Burn Pneumonia

doi: 10.3390/ebj7010001

Figure Lengend Snippet: Elevated plasma Angiopoietin-2 and downstream pulmonary mediators in a murine model of burn injury. Heparinized plasma was collected one day after burn or sham injury and evaluated for Ang-2 via ELISA. Dark blue dots represent mice that received sham injury, red dots represent mice that received burn injury. ( A ) Plasma Ang-2 was significantly increased in burn mice compared to sham. ( B ) Representative IHC of Ang-2 in sham and burn lungs. ( C – H ) Lung tissue was flash frozen one day after burn injury and then isolated for RNA, and RT-qPCR was performed with GAPDH as housekeeping control. ( C ) RT-qPCR expression of Tek, the transcript for the TIE2 receptor, was significantly elevated in burn lungs compared to sham ( p < 0.01). ( D ) Tek expression positively correlated with plasma Ang-2 levels (simple-linear regression, R 2 = 0.5655, p < 0.01). ( E ) Tnip2, the transcript of ABIN2, and ( F ) Pik3r1, a subunit of PI3K, were both significantly upregulated in the lungs following burn injury. ( G ) ICAM-1 and ( H ) VCAM-1 were also both significantly upregulated in the lungs following burn injury. For two-group analysis, significance was analyzed by unpaired t -test, n = 6–8 animals per group, ** p < 0.01, **** p < 0.0001. ( I – L ) Lung expression of Tnip2, Pik3r1, ICAM-1, and VCAM-1 all positively correlated with plasma Ang-2 levels (simple-linear regression, p < 0.001).

Article Snippet: Plasma Angiopoietin-2 levels were determined using Mouse & Rat Angiopoietin-2 Quantikine ELISA Kit (R&D Systems Cat. MANG20) using a 1:40 dilution of plasma in technical duplicate and reported as ng of Ang-2/mL of plasma.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Isolation, Quantitative RT-PCR, Control, Expressing

Journal: European Burn Journal

Article Title: The Role of Angiopoietin-2 in Post-Burn Pneumonia

doi: 10.3390/ebj7010001

Figure Lengend Snippet: Pulmonary neutrophil infiltration correlates with plasma Angiopoietin-2. Dark blue dots represent mice that received sham injury, red dots represent mice that received burn injury. ( A ) Representative H&E of lungs of sham and burn mice. ( B ) Neutrophils were counted in 6 high-powered fields in 3 mice each of sham and burn ( p < 0.0001). ( C ) Lung RT-qPCR of chemokine Cxcl1 was elevated in burn compared to sham ( p < 0.05) and ( H ) positively correlated with plasma Ang-2 ( p < 0.01). ( D – G ) Neutrophil markers ( D ) Lcn2 ( p < 0.01), ( E ) Ly6g ( p = 0.0737), ( F ) s100a8 ( p < 0.05), and ( G ) s100a9 ( p < 0.05) were elevated in burn compared to sham and ( I – L ) all positively correlated with plasma Ang-2 levels ( p < 0.05). For two-group analysis, significance was analyzed by unpaired t -test and correlation assessed by simple-linear regression, * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Plasma Angiopoietin-2 levels were determined using Mouse & Rat Angiopoietin-2 Quantikine ELISA Kit (R&D Systems Cat. MANG20) using a 1:40 dilution of plasma in technical duplicate and reported as ng of Ang-2/mL of plasma.

Techniques: Clinical Proteomics, Quantitative RT-PCR