Journal: PLOS Biology

Article Title: The suprachiasmatic nucleus regulates brown fat thermogenesis in male mice through an adrenergic receptor ADRB3-S100B signaling pathway

doi: 10.1371/journal.pbio.3003534

Figure Lengend Snippet: (A) Volcano plot showing differentially expressed genes (DEGs) in interscapular BAT from sham and SCNx mice under TRF-STE. DEGs were defined by fold change > 2, p < 0.01. Commonly upregulated genes are highlighted in red and downregulated genes in green at both ZT4 and ZT16. n = 3 per group. (B) IPA of DEGs at ZT4 and ZT16. Pathways with significant enrichment ( p < 0.01) are shown, with activation z-scores indicated for each pathway. (C) DAVID pathway analysis of S100 family-associated DEGs. Enriched KEGG pathways are shown in yellow, and Gene Ontology biological processes (BP) are shown in gray. (D and E) Correlation analysis of RNA-seq-derived S100b expression with cell proliferation markers ( D ) and senescence-related genes ( E ) in sham and SCNx mice. Pearson correlation coefficients and p -values are shown. (F and G) Cell proliferation analysis via EdU incorporation. Representative images showing EdU incorporation in interscapular BAT from sham and SCNx mice at ZT4 and ZT16 under Ad-STE and TRE-STE with 4°C and 30°C as control (F) . Quantification of EdU + nuclei as a percentage of total (Hoechst-stained) nuclei (G) . n = 5 per group. (H–K) Assessment of S100B-induced proliferation in PDGFRα + preadipocytes. PDGFRα-positive stromal vascular fraction (SVF) cells (H) , representative images of EdU-positive cells treated with recombinant S100B (I) , statistical analysis of EdU + nuclei (J) , and cell growth curves of control vs. S100B-treated preadipocytes (K) . (L) Expression of differentiation-related genes during SVF from interscapular BAT induced to differentiate into mature adipocytes. n = 6 per group. (M) Representative β-galactosidase staining images showing senescent cell abundance in interscapular BAT from sham and SCNx mice under TRF-STE. Data are presented as mean ± SD. Statistical significance was determined using unpaired two-tailed Student t test ( G , J, and L ), Pearson correlation analysis ( D and E ), and two-way ANOVA with Sidak’S multiple comparisons test (K) . * p < 0.05, ** p < 0.01 and **** p < 0.0001. Scale bars, 100 μm (F and M) , 50 μm (I) , 20 μm ( H and magnified view of I ). The data underlying the graphs shown in the figure can be found in

Article Snippet: Recombinant Mouse S100B , Targetmol , Cat# TMPJ-00990.

Techniques: Activation Assay, RNA Sequencing, Derivative Assay, Expressing, Control, Staining, Recombinant, Two Tailed Test

Journal: PLOS Biology

Article Title: The suprachiasmatic nucleus regulates brown fat thermogenesis in male mice through an adrenergic receptor ADRB3-S100B signaling pathway

doi: 10.1371/journal.pbio.3003534

Figure Lengend Snippet: (A) S100b expression levels under various nutritional states (fasting, HFD, and TRF) and environmental temperatures (4°C, 21°C, and 30°C). n = 4 per group. (B) S100b and Ucp1 expression levels in indicated temperatures. n = 4 per group. (C) Serum S100B levels were measured by ELISA in sham and SCNx mice under Ad-STE and TRF-STE at ZT4 and ZT16. n = 4 per group. (D and E) Serum biochemical assays showing levels of TG ( D ) and NEFA ( E ) in sham and SCNx mice under Ad-STE ( n = 6 per group). (F) Representative immunohistochemistry images of S100B protein in interscapular BAT from sham and SCNx mice under Ad-STE and TRF-STE conditions. Scale bars, 50μm. (G and H) Western blot analysis ( G ) and densitometric quantification ( H ) of S100B, p21, and CCND1 protein levels in interscapular BAT from sham and SCNx mice at ZT4 and ZT16. n = 4 per group. Red arrows indicate quantified bands. (I) Relative mRNA levels of indicated genes in interscapular BAT from sham and SCNx BAT following ADRB3 antagonist with SR59230A. n = 6 per group. SR: SR59230A. (J and K) Structural prediction of the human ADRB3-S100B complex using AlphaFold2/ColabFold. The human ADRB3/S100B complex model ( J ) was predicted by ColabFold and colored in blue, cyan, yellow, and orange according to different prediction confidence (pLDDT, predicted local distance difference test). Surface electrostatic representation showing ADRB3 (deep teal) and S100B (orange) with positive and negative charges indicated in blue and red, respectively (K) . ( L ) coIP of ADRB3 and S100B in HEK293T cells. (M and N) Western blot ( M ) and densitometry analysis ( N ) of ADRB3 protein levels in interscapular BAT following S100b knockdown or overexpression in vivo. n = 4 per group. Red arrows indicate quantified bands. (O and P) ECAR analysis ( O ) and quantification of basal glycolysis and glycolytic capacity ( P ) in primary preadipocytes treated with S100B and/or β3-agonist CL-316243. Glucose, oligomycin, and 2-DG were sequentially injected. Data are presented as mean ± SEM. Control, CL, 0.1 µM S100B and 0.2 µM S100B: n = 6; 0.1 µM S100B + CL and 0.2 µM S100B+CL: n = 3. (Q) Schematic model illustrating the proposed ADRB3-S100B signaling axis. SCN lesioning enhances SNS activity, thereby promoting ADRB3 signaling and upregulating S100B expression. S100B, in turn, increases ADRB3 sensitivity, establishing a positive feedback loop that sustains thermogenesis and stimulates preadipocyte proliferation. Created in BioRender.com. Unless otherwise indicated, data are presented as mean ± SD. NS: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 by unpaired two-tailed Student t test ( A – E , H , I , N, and P ). The data underlying the graphs shown in the figure can be found in S1 Source Data. Raw blot images can be found in S1 Raw Images.

Article Snippet: Recombinant Mouse S100B , Targetmol , Cat# TMPJ-00990.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Western Blot, Structural Proteomics, Knockdown, Over Expression, In Vivo, Injection, Control, Activity Assay, Two Tailed Test

Journal: PLOS Biology

Article Title: The suprachiasmatic nucleus regulates brown fat thermogenesis in male mice through an adrenergic receptor ADRB3-S100B signaling pathway

doi: 10.1371/journal.pbio.3003534

Figure Lengend Snippet: (A) Schematic of AAV-mediated S100b knockdown or scramble control delivery into interscapular BAT of SCNx mice. (B) Relative mRNA expression of the indicated genes in interscapular BAT from SCNx mice under TRF-STE with scramble or S100b targeting shRNA. n = 6 per group. (C and D) Western blot analysis of S100B and p21 ( C ) and corresponding densitometry quantification ( D ) in interscapular BAT from SCNx mice under TRF-STE ( n = 4 per group). (E and F) EdU incorporation analysis in interscapular BAT from SCNx mice following S100b knockdown or scramble control. Representative images ( E ) and quantification of EdU-positive nuclei relative to total Hoechst-positive nuclei (F) . (G) Body temperature profiles of SCNx mice with S100b knockdown or scramble control under Ad-STE and TRF-STE (scramble: n = 6, S100b knockdown: n = 5). Data presented as mean ± SEM. Corresponding interscapular BAT and tail temperature data are shown in . (H) Schematic illustrating AAV-mediated overexpression of S100b (AAV-DIO- S100b + AAV-CAG-Cre) or EGFP control in interscapular BAT of WT mice. (I) Relative mRNA levels of indicated genes in interscapular BAT from EGFP or S100b overexpressing mice under TRF-STE ( n = 6 per group). (J and K) Western blot analysis of S100B and p21 proteins ( J ) and corresponding densitometry ( K ) from interscapular BAT of EGFP or S100b -overexpressing mice ( n = 4 per group). (L and M) EdU incorporation analysis in interscapular BAT from EGFP and S100b -overexpressing mice. Representative images ( L ) and quantification of EdU + cells (M) . n = 6 per group. (N) Body temperature responses under Ad-STE and TRF-STE in WT mice with EGFP or S100b overexpression in interscapular BAT. Data presented as mean ± SEM. n = 6 per group. See for interscapular BAT and tail temperature profiles. Unless otherwise indicated, data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. Significance determined by unpaired two-tailed Student t test ( B , D , F , I , K, and M ) or two-way ANOVA with Sidak’S multiple comparisons test (G, N) . Scale bars, 50 μm (E, L) , 20 μm (magnified view of E , L ). Schematic ( A , H ) created in BioRender.com. The data underlying the graphs shown in the figure can be found in S1 Source Data. Raw blot images can be found in S1 Raw Images.

Article Snippet: Recombinant Mouse S100B , Targetmol , Cat# TMPJ-00990.

Techniques: Knockdown, Control, Expressing, shRNA, Western Blot, Over Expression, Two Tailed Test

Journal: PLOS Biology

Article Title: The suprachiasmatic nucleus regulates brown fat thermogenesis in male mice through an adrenergic receptor ADRB3-S100B signaling pathway

doi: 10.1371/journal.pbio.3003534

Figure Lengend Snippet: (A) LL exposure paradigm is used to disrupt SCN rhythmicity. Arrhythmic mice were selected for subsequent analysis . (B) Relative mRNA expression of indicated genes in interscapular BAT from WT mice under light/dark cycle (LD) or LL during TRF-STE ( n = 4 per group). (C and D) Western blot analysis ( C ) and densitometry ( D ) of S100B and p21 proteins in interscapular BAT from mice under LD or LL conditions ( n = 4 per group). (E and F) EdU staining in interscapular BAT from mice under LD or LL. Representative images ( E ) and quantification of EdU + cells ( F ) normalized to Hoechst staining. (G) Body temperature profiles of WT mice under LD or LL conditions during TRF-STE. Data presented as mean ± SEM. n = 4 per group. See for additional temperature metrics. (H) Working model illustrating the SCN-ADRB3-S100B axis in BAT during TRF conducted at ZT16-ZT20 in a subthermoneutral environment, the SCN regulates BAT thermogenic plasticity via SNS output. SCN lesioning enhances ADRB3 signaling and S100B expression, which together form a positive feedback loop that amplifies β3-adrenergic sensitivity, promotes preadipocyte proliferation, suppresses senescence, and sustains glucose-driven thermogenesis. Unless otherwise indicated, data are presented as mean ± SD. * p < 0.05, ** p < 0.01 and **** p < 0.0001. Significance determined by unpaired two-tailed Student t test ( B , D, and F ) or two-way ANOVA with Sidak’S multiple comparisons test (G) . Scale bars, 50 μm (E) , 20 μm (magnified view of E ). Schematic ( A ) and working model ( H ) created in BioRender.com. The data underlying the graphs shown in the figure can be found in S1 Source Data. Raw blot images can be found in S1 Raw Images.

Article Snippet: Recombinant Mouse S100B , Targetmol , Cat# TMPJ-00990.

Techniques: Expressing, Western Blot, Staining, Two Tailed Test

Journal: Experimental and Therapeutic Medicine

Article Title: The protective and hemodynamic effects of dexmedetomidine on hypertensive cerebral hemorrhage patients in the perioperative period

doi: 10.3892/etm.2016.3711

Figure Lengend Snippet: Levels of NSE and S100β.

Article Snippet: The catecholamine (Rocky Mountain Diagnostics, Colorado Springs, CO, USA), neuron-specific enolase (NSE) and S100β (S100β protein) were estimated using ELISA as per instructions the manufacturer's (Rocky Mountain Diagnostics).

Techniques:

Journal: Journal of Veterinary Internal Medicine

Article Title: Time course and prognostic value of serum GFAP, pNFH, and S100β concentrations in dogs with complete spinal cord injury because of intervertebral disc extrusion

doi: 10.1111/jvim.15439

Figure Lengend Snippet: Scatter plots of A, GFAP; B, s100β; and C, pNFH serum concentrations in 31 dogs over 14 days after onset of nonambulatory status. Dogs are identified according to whether or not they are walking at 6 months, with white squares representing dogs that did not recover and solid black circles representing dogs that did recover. GFAP, glial fibrillary acidic protein; pNFH, phosphorylated neurofilament heavy chain

Article Snippet: For S100β, an ELISA detection kit for Canine S100β Protein from MyBiosource (San Diego, California) was used.

Techniques:

Journal: Journal of Veterinary Internal Medicine

Article Title: Time course and prognostic value of serum GFAP, pNFH, and S100β concentrations in dogs with complete spinal cord injury because of intervertebral disc extrusion

doi: 10.1111/jvim.15439

Figure Lengend Snippet: Serum concentrations of GFAP, pNFH, and S100β expressed as median and range in 31 dogs categorized as walking at 6 months, yes or no and grouped as time of presentation as well as time of sampling after onset of nonambulatory status (1, 2, or 3 days). The number of samples analyzed at each time point is listed beneath the range

Article Snippet: For S100β, an ELISA detection kit for Canine S100β Protein from MyBiosource (San Diego, California) was used.

Techniques: Sampling

Journal: Journal of Veterinary Internal Medicine

Article Title: Time course and prognostic value of serum GFAP, pNFH, and S100β concentrations in dogs with complete spinal cord injury because of intervertebral disc extrusion

doi: 10.1111/jvim.15439

Figure Lengend Snippet: The estimated probability of response (ambulatory yes or no) at 6 months with the observed data for A, log average of GFAP and pNFH; B, log average of GFAP and S100β; and C, log average of pNFH and S100β overlaid. The heat map scale of probability of response ranges from 0, indicating recovery of walking, to 1, indicating no recovery of walking. White squares are dogs that did not recover; black circles are dogs that did recover. GFAP, glial fibrillary acidic protein; pNFH, phosphorylated neurofilament heavy chain

Article Snippet: For S100β, an ELISA detection kit for Canine S100β Protein from MyBiosource (San Diego, California) was used.

Techniques: