Journal: International Journal of Molecular Sciences

Article Title: Distinct Plasma Immune Profile in ALS Implicates sTNFR-II in pAMPK/Leptin Homeostasis

doi: 10.3390/ijms24065065

Figure Lengend Snippet: Plasma levels of leptin, CCL16 and sTNF-RII measured by ELISA. ( a ) concentration (ng/mL) of leptin measured by ELISA in healthy controls (38) and sALS (51). ( b , d ) Concentration (ng/mL) of leptin ( b ), CCL16 ( c ) and sTNF-RII ( d ) measured by ELISA in slow (40) and fast (11) progressing ALS patients. ( e , f ) Concentration (ng/mL) of leptin in sALS and controls sub-grouped by sex ( e ) and by sex and rate of progression ( f ) (Data are mean ± SEM * p ≤ 0.05, ** p < 0.01, **** p < 0.0001, NS, not significant).

Article Snippet: Cells were also treated with different concentrations of recombinant CCL16 (R&D systems, Minneapolis, MN, USA, # 802-HC-025) or recombinant sTNF-RII (MyBioSource, MBS343136, London, UK).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Distinct Plasma Immune Profile in ALS Implicates sTNFR-II in pAMPK/Leptin Homeostasis

doi: 10.3390/ijms24065065

Figure Lengend Snippet: Exposure to plasma from sALS reduces leptin production in human adipocytes. ( a ) Oil red O coloration demonstrating lipid droplet accumulation (red) in mature adipocytes. ( b ) Human adipocytes were treated for 12 h with 1:100 diluted mixed plasma samples from healthy controls (12), slow progressors (12) and fast progressors (6). The supernatant was collected, and leptin levels were measured by ELISA (pg/mL) and normalized with plasma sample levels of leptin. ( c ) Leptin levels (pg/mL) secreted by mature adipocytes after plasma samples treatment. The adipocytes were pretreated with 1 μM of mTOR inhibitor (PP242) or 10 μM of AMPK inhibitor (compound C) for 1h. ( d ) Proteins from treated adipocytes ( c ) were extracted using RIPA buffer. Graph exhibits pAMPK/GAPDH fold change observed by immunoblotting with or without 10 μM AMPK inhibitor pretreatment. ( e , f ) Leptin levels (pg/mL) secreted by mature adipocytes after insulin and sTNF-RII treatment ( e ) and insulin and CCL16 treatment ( f ). ( g ) Immunoblotting showing pAMPK levels in adipocytes treated with increasing concentration of sTNF-RII. ( h ) Quantification of immunoblotting in ( g ) showing levels of pAMPK/GAPDH fold change in adipocytes treated with sTNF-RII ((Data are mean ± SEM * p ≤ 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001) n = 3, two-way ANOVA). NS, not significant.

Article Snippet: Cells were also treated with different concentrations of recombinant CCL16 (R&D systems, Minneapolis, MN, USA, # 802-HC-025) or recombinant sTNF-RII (MyBioSource, MBS343136, London, UK).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Western Blot, Concentration Assay