Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: S100β induces neurite loss (A and B) Low- and high-magnification views of brain sections from Thy1-YFP mice harvested 3 days after S100β injection. Scale bar: 500 μm. In (A), the arrow shows the injection site, and the rectangle shows the area of higher-magnification views shown in (B). In (B), neuronal cell nuclei are identified by NeuN immunostaining (magenta). Scale bar: 10 μm. (C) Quantification of neuronal cell body density. (D and E) YFP neurite length and area, expressed relative to saline-injected controls. n = 4; *p < 0.05 and **p < 0.01 vs. saline controls by one-way ANOVA with Dunnett’s test. All data are shown as mean ± SEM.

Article Snippet: Recombinant mouse S100β , , Novus Biologicals , Cat# NBP2-53070.

Techniques: Injection, Immunostaining, Saline

Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: S100β induces cofilactin rod (CAR) formation and neurite loss (A) Diagram shows the locations of intracortical injections and peri-injection region imaged. (B) CAR formation identified by immunostaining for cofilin-1 aggregates (red) in WT mice after saline or 50 ng S100β injections. Neurites are identified by neurofilament-H (NF-H; green). NF-H integrity is lost at sites of CAR formation. Scale bar: 10 μm. (C–E) CAR density expressed as percentage of total neurite area. (F) Neurite loss 1, 3, and 7 days after saline or 50 ng S100β injections as assessed by immunostaining for NF-H (green), with neuronal cell nuclei identified by NeuN (magenta). Scale bar: 10 μm. (G–I) NF-H+ neurite length per neuronal nucleus in the S100β-injected mice. n = 4; **p < 0.01 by Student’s t test. All data are shown as mean ± SEM.

Article Snippet: Recombinant mouse S100β , , Novus Biologicals , Cat# NBP2-53070.

Techniques: Injection, Immunostaining, Saline

Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: CAR formation is attenuated in both cofilin-1 hemizygous and p47 phox -deficient mice (A) CAR formation after saline or 50 ng S100β injections in WT, COF −/+ , and p47 phox−/− mice identified by immunostaining for cofilin-1 aggregates (red). Neurites are identified by NF-H (green). Scale bar: 10 μm. (B–D) CAR density expressed as percentage of total neurite area. (E) Neurite loss after saline or 50 ng S100β injections as assessed by immunostaining for NF-H (green), with neuronal cell nuclei identified by NeuN (magenta). Scale bar: 20 μm. (F–H) Neurite length per neuronal nucleus in the S100β-injected mice relative to saline-injected mice of each genotype. n = 4; *p < 0.05 and **p < 0.01 vs. WT mice by one-way ANOVA with Dunnett’s test. All data are shown as mean ± SEM.

Article Snippet: Recombinant mouse S100β , , Novus Biologicals , Cat# NBP2-53070.

Techniques: Saline, Immunostaining, Injection

Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: S100β-induced neurite loss and motor impairment are attenuated in cofilin-1 hemizygous and p47 phox -deficient mice (A) Schematic timeline of behavioral assessments. (B) Performance on the corner test assessed by quantifying the percentage of left turns. n = 6 for each genotype. *p < 0.05 and **p < 0.01 vs. WT by one-way ANOVA with Dunnett’s test. (C) Iba-1 immunostaining shows microglial/macrophage (M/M) activation 1 day after injection with 50 ng S100β. Insets show magnified views. Responses to S100β injections were similar in the three mouse genotypes. Scale bars: 10 μm. (D–F) Quantification of M/M responses to S100β. n = 4; *p < 0.05 and **p < 0.01 vs. WT mice by one-way ANOVA with Dunnett’s test. All data are shown as mean ± SEM.

Article Snippet: Recombinant mouse S100β , , Novus Biologicals , Cat# NBP2-53070.

Techniques: Immunostaining, Activation Assay, Injection

Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: Neurite loss in neuron-glia co-cultures is attenuated by neuronal cofilin-1 hemizygosity and glial p47 phox deficiency (A) Photomicrographs of co-cultures immunostained for MAP-2 (green) and NeuN (blue). Scale bar: 20 μm. (B–D) Neurite length assessed at 4, 24, and 48 h of 50 ng/mL S100β incubation, expressed relative to control wells of the respective co-culture type. n = 4; *p < 0.05 and **p < 0.01 by one-way ANOVA with Dunnett’s test. (E) Mechanism proposed for inflammation-induced neurite degeneration. In response to pro-inflammatory stimuli, brain microglia and infiltrating macrophages upregulate superoxide production by NADPH oxidase. Resulting oxidative stress in nearby neurites leads to formation of CARs. Persistence of the CARs causes neurite degeneration, which can occur in the absence of parental neuron death. This process is attenuated in p47 phox−/− mice, which cannot form an active NADPH oxidase-2 complex, and in cofilin hemizygous ( COF −/+ ) mice, which have reduced propensity to form CARs. All data are shown as mean ± SEM.

Article Snippet: Recombinant mouse S100β , , Novus Biologicals , Cat# NBP2-53070.

Techniques: Incubation, Control, Co-Culture Assay

Journal: Cell reports

Article Title: Cofilactin rod formation mediates inflammation-induced neurite degeneration

doi: 10.1016/j.celrep.2024.113914

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Recombinant mouse S100β , , Novus Biologicals , Cat# NBP2-53070.

Techniques: Affinity Purification, Recombinant, Membrane, Software

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: For EdU-labeled cell proliferation of SVF, 0.1 μM recombinant S100B protein (TMPJ-00990, Targetmol USA) or saline and 10 μM EdU were added to the medium.

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: For EdU-labeled cell proliferation of SVF, 0.1 μM recombinant S100B protein (TMPJ-00990, Targetmol USA) or saline and 10 μM EdU were added to the medium.

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: For EdU-labeled cell proliferation of SVF, 0.1 μM recombinant S100B protein (TMPJ-00990, Targetmol USA) or saline and 10 μM EdU were added to the medium.

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: For EdU-labeled cell proliferation of SVF, 0.1 μM recombinant S100B protein (TMPJ-00990, Targetmol USA) or saline and 10 μM EdU were added to the medium.

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

Journal: International journal of oncology

Article Title: S100B expression in breast cancer as a predictive marker for cancer metastasis.

doi: 10.3892/ijo.2017.4226

Figure Lengend Snippet: Figure 1. Expression levels of S100B in normal and tumor tissue. The expression of S100B was assessed from the Oncomine database. (A) TCGA breast dataset; the bars are labeled as follows: 1, breast (normal, n=61); 2, invasive ductal breast carcinoma (n=389); 3, invasive breast carcinoma (n=76); mixed lobular and ductal breast carcinoma (n=7); 5, invasive lobular breast carcinoma (n=36). (B) The Ma et al breast dataset (19); the bars are labeled as follows: 1, breast carcinoma (n=14); 2, ductal breast carcinoma in situ (n=9); 3, invasive ductal breast carcinoma (n=9). Boxplot shows the z-score of S100B expression according to the raw data from Oncomine. Error bars represent standard deviation (SD). The statistically significant differences between each group were examined with one-way ANOVA (***P<0.001). (C) S100B expression in different types of breast cancer examined via the GOBO database. (D) S100B expres- sion was examined in estrogen receptor (ER)-negative (ER-neg) and ER-positive (ER-pos) breast cancer using the GOBO database. The number above each bar indicates the sample size in each group. (E) mRNA expression of S100B in breast cancer cell lines. The error bars represent SD.

Article Snippet: After scratching, cell debris was removed by washing twice with phosphate-buffered saline (PBS) and the cells were then incubated in serum-free L-15 medium containing 0, 0.1 and 1 nM recombinant human S100B protein (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Labeling, In Situ, Standard Deviation

Journal: International journal of oncology

Article Title: S100B expression in breast cancer as a predictive marker for cancer metastasis.

doi: 10.3892/ijo.2017.4226

Figure Lengend Snippet: Figure 2. S100B treatment inhibits cell migration. Transwell migration assay in (A) MDA-MB-231 and (B) Hs578T and (C) MCF-7 cells. Images and quanti- fication results are shown. Images of wound healing assay in (D) MDA-MB-231 and (E) MCF-7 cells. The quantification of wound healing assay in both cells is shown in the lower panel in the images. Error bars represent SD (one-way ANOVA; **p<0.01 and ***p<0.001).

Article Snippet: After scratching, cell debris was removed by washing twice with phosphate-buffered saline (PBS) and the cells were then incubated in serum-free L-15 medium containing 0, 0.1 and 1 nM recombinant human S100B protein (R&D Systems, Minneapolis, MN, USA).

Techniques: Migration, Transwell Migration Assay, Wound Healing Assay

Journal: International journal of oncology

Article Title: S100B expression in breast cancer as a predictive marker for cancer metastasis.

doi: 10.3892/ijo.2017.4226

Figure Lengend Snippet: Figure 3. S100B treatment induces mesenchymal-epithelial transition (MET) in MDA-MB-231 cells. (A and B) Western blot analysis was used to examine the protein expression levels of MET signaling pathway-related molecules. Band intensity was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The relative value of the control (0 nM) was set to 1. Error bars represent SD (t-test; *p<0.05 and **p<0.01).

Article Snippet: After scratching, cell debris was removed by washing twice with phosphate-buffered saline (PBS) and the cells were then incubated in serum-free L-15 medium containing 0, 0.1 and 1 nM recombinant human S100B protein (R&D Systems, Minneapolis, MN, USA).

Techniques: Western Blot, Expressing, Control

Journal: International journal of oncology

Article Title: S100B expression in breast cancer as a predictive marker for cancer metastasis.

doi: 10.3892/ijo.2017.4226

Figure Lengend Snippet: Figure 4. Evaluation of the association between S100B expression and survival curve. (A) S100B expression in the low-risk (green) and high-risk (red) group. The box plot is adapted from the SurvExpress database and shows S100B expression in patients. (B) Kaplan-Meier distant recurrence curve was created using the SurvExpress database to analyze the sample from a GEO dataset (GSE9893). The low-risk (n=67) and high-risk (n=67) groups are shown in green and red, respectively. (C) Analyzing the overall survival rate in all breast cancer patients, (D) estrogen receptor (ER)-positive breast cancer patients, and (E) ER-negative breast cancer patients in ‘breast invasive carcinoma TCGA’ from the SurvExpress database. The green and red lines indicate a high and low S100B expression, respectively. Evaluation of the distant metastasis-free survival (DMFS) curve comparing the patients with a high (red) and low (black) S100B expression in (F) all breast cancer patients, (G) ER-positive breast cancer patients, (H) ER-negative breast cancer patients and (I) endocrine therapy-treated breast cancer patients using the KM plotter database. The hazard ratio (HR) and log-rank P-values are shown in each panel.

Article Snippet: After scratching, cell debris was removed by washing twice with phosphate-buffered saline (PBS) and the cells were then incubated in serum-free L-15 medium containing 0, 0.1 and 1 nM recombinant human S100B protein (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing

Journal: International journal of oncology

Article Title: S100B expression in breast cancer as a predictive marker for cancer metastasis.

doi: 10.3892/ijo.2017.4226

Figure Lengend Snippet: Figure 5. Evaluation of whether p53 expression in human breast cancer is associated with S100B expression and patient survival. (A) Expression of S100B in patients with or without p53 mutation. Raw data were adapted from the TCGA breast dataset. Distant metastasis-free survival (DMFS) analysis in breast cancer patients with (B) p53 mutation and (C) p53 wild-type. The hazard ratio (HR) and log-rank P-value are shown in each panel.

Article Snippet: After scratching, cell debris was removed by washing twice with phosphate-buffered saline (PBS) and the cells were then incubated in serum-free L-15 medium containing 0, 0.1 and 1 nM recombinant human S100B protein (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Mutagenesis

Journal: International journal of oncology

Article Title: S100B expression in breast cancer as a predictive marker for cancer metastasis.

doi: 10.3892/ijo.2017.4226

Figure Lengend Snippet: Figure 6. Summary of the S100B-mediated effects in breast cancer. Patients with breast cancer expressing high levels of S100B exhibited a good prognosis and a low metastatic rate. When the breast cancer cells are treated with S100B, the migration ability was inhibited and the epithelial phenotype was induced.

Article Snippet: After scratching, cell debris was removed by washing twice with phosphate-buffered saline (PBS) and the cells were then incubated in serum-free L-15 medium containing 0, 0.1 and 1 nM recombinant human S100B protein (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Migration