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: Frontiers in Cellular and Infection Microbiology

Article Title: S100B Inhibition Attenuates Intestinal Damage and Diarrhea Severity During Clostridioides difficile Infection by Modulating Inflammatory Response

doi: 10.3389/fcimb.2021.739874

Figure Lengend Snippet: S100B inhibition upregulates anti-inflammatory ( SOCS2 ) and antiapoptotic mediators ( Bcl2 ) and downregulates inflammatory mediators during CDI. (A, B) Heat map of the cDNA microarray analysis of colonic tissues from non-pretreated C. difficile -infected (CDI) and pentamidine-pretreated C. difficile- infected (CDI+pentamidine) mice at day 3 p.i. Expression of the genes is normalized to median of the control, log 2 scale. TaqMan qPCR analysis of (C) proinflammatory (D) chemokine, (E) cellular recruitment ( SELP ), (F) anti-inflammatory SOCS2 , and (G) antiapoptotic Bcl2 mediators. Data are mean ± s.e.m. (C–E) # p < 0.05, *p < 0.01, **p < 0.001. ANOVA followed by Turkey test was used.

Article Snippet: The sections were then incubated with an S100B antibody (NBP2-54426, Novus Biologicals, 1:1,000) overnight.

Techniques: Inhibition, Microarray, Infection, Expressing, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: S100B Inhibition Attenuates Intestinal Damage and Diarrhea Severity During Clostridioides difficile Infection by Modulating Inflammatory Response

doi: 10.3389/fcimb.2021.739874

Figure Lengend Snippet: C. difficile infection increases S100B in fecal samples and in colon from humans and mice. (A) Representative immunohistochemical images of S100Bexpression in human colonic biopsies with active C. difficile infection (CDI) and healthy subjects (Control). Increased S100B expression (arrowhead or rectangle) was found in colonic mucosal (center panel, red arrowhead), submucosal (center panel, black arrowhead), and myenteric plexus (right panel, black rectangle). Scale bars, 200 (left panels), 100 (center panels), and 50 (right panels) μm. Left panels are showing colon tissues from control and CDI patients in a low magnification. (B) Quantification of percentage (mean ± s.e.m.) of S100B-immunopositive area in colons from human control and C. difficile- infected subjects in 15–20 microscope fields per sample (n = 4 subjects per group). Unpaired two-tailed Student t test. (C) S100B levels in fecal samples of patients with diarrhea caused by CDI (n = 53) and non-CDI (n = 27) evaluated by ELISA. Data are median ± s.d. Two-tailed non-parametric Mann–Whitney U-test. (D) Schematic diagram of CDI experimental model in mice. (E) Representative Western blot (WB) bands of S100B and α-tubulin in colonic tissues from mice infected with C. difficile (CDI group) and non-infected (control group) at days 1 and 3 postinfection (p.i.). (F) WB analysis of S100B (mean ± s.e.m.) in colonic tissues from CDI and control group (n = 3 mice per group). α-Tubulin was used to normalize the levels of S100B. Unpaired two-tailed Student’s t test. (G) Representative immunohistochemical images of S100B immunostaining in colonic tissues of mice with CDI and control (non-infected mice). (H) Quantification of percentage (mean ± s.e.m.) of S100B-immunopositive area in colon from mice with CDI and control (non-infected mice) (n = 5 mice per group). Unpaired two-tailed Student’s t test.

Article Snippet: The sections were then incubated with an S100B antibody (NBP2-54426, Novus Biologicals, 1:1,000) overnight.

Techniques: Infection, Immunohistochemical staining, Control, Expressing, Microscopy, Two Tailed Test, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Western Blot, Immunostaining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: S100B Inhibition Attenuates Intestinal Damage and Diarrhea Severity During Clostridioides difficile Infection by Modulating Inflammatory Response

doi: 10.3389/fcimb.2021.739874

Figure Lengend Snippet: S100B inhibition decreases diarrhea severity, intestinal damage, and neutrophil recruitment during CDI in mice. Mice were infected with 10 5 vegetative C. difficile (VPI10463 strain) and treated with a S100B inhibitor, pentamidine (40 mg/kg, i.p., once daily for 3 days, CDI+pentamidine group) or non-treated (CDI group). (A) Quantification of C. difficile shedding (mean ± s.e.m.) in stools by amplifying the tcdB gene by qPCR. Unpaired two-tailed Student’s t test. (B) Diarrhea score (median ± s.d) of CDI and CDI+pentamidine mice. Two-tailed non-parametric Mann–Whitney U-test. (C) Representative H&E stains of cecal and colonic tissues collected from CDI and CDI+pentamidine mice at day 3 postinfection (p.i.). CDI promotes damage of colonic and cecal epithelium (black arrow), edema (green arrow), and inflammatory cell infiltration (red arrow) in mice infected by C. difficile (CDI group). Scale bars, 100 µm. (D) Histopathologic score (median, 0-no damage, and 9-intense damage) based on epithelial damage, submucosal edema, and infiltration of inflammatory cells. Two-tailed non-parametric Mann–Whitney U-test. (E) MPO levels (mean ± s.e.m.) in cecum content, cecum, and colon samples from CDI and CDI+pentamidine mice at day 3 p.i. measured by ELISA. Unpaired two-tailed Student’s t test.

Article Snippet: The sections were then incubated with an S100B antibody (NBP2-54426, Novus Biologicals, 1:1,000) overnight.

Techniques: Inhibition, Infection, Two Tailed Test, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: S100B Inhibition Attenuates Intestinal Damage and Diarrhea Severity During Clostridioides difficile Infection by Modulating Inflammatory Response

doi: 10.3389/fcimb.2021.739874

Figure Lengend Snippet: S100B modulates the release of pro-inflammatory mediators and tissue repair cytokines during CDI in mice. Levels of (A) IL-1β, (B) IL-18, (C) IL-6, (D) GMCSF, (E) TNF-α, (F) IL-17, (G) IL-23, (H) IL-2, (I) IL-33, and (J) IL-22 in colonic tissues from uninfected (control), uninfected receiving pentamidine (40 mg/kg, pentamidine), non-pretreated C. difficile -infected (CDI), and pentamidine-pretreated C. difficile- infected (CDI+pentamidine) mice at day 3 p.i. were measured by ELISA. Data are mean ± s.e.m. **ANOVA followed by Turkey test was used. *Unpaired two-tailed Student’s t test.

Article Snippet: The sections were then incubated with an S100B antibody (NBP2-54426, Novus Biologicals, 1:1,000) overnight.

Techniques: Control, Infection, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: S100B Inhibition Attenuates Intestinal Damage and Diarrhea Severity During Clostridioides difficile Infection by Modulating Inflammatory Response

doi: 10.3389/fcimb.2021.739874

Figure Lengend Snippet: TcdA and TcdB increase S100B-dependent upregulation of IL-6 expression in enteroglial cells (EGC/PK060399). (A, B) Levels of S100B (mean ± s.e.m) released by ELISA, (C, D) S100B , and (E, F) IL-6 gene expression (mean ± s.e.m) by qPCR in enteroglial cells (EGC/PK060399) challenged with (A, C, E) TcdA and (B, D, F) TcdB (n = 4) with TcdA and TcdB (n = 4). (A–F) Cells receiving only supplemented DMEM were applied as a control. (A–F) ANOVA followed by Sidak’s multiple-comparison test was used.

Article Snippet: The sections were then incubated with an S100B antibody (NBP2-54426, Novus Biologicals, 1:1,000) overnight.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Gene Expression, Control, Comparison

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: S100B Inhibition Attenuates Intestinal Damage and Diarrhea Severity During Clostridioides difficile Infection by Modulating Inflammatory Response

doi: 10.3389/fcimb.2021.739874

Figure Lengend Snippet: Blockade of the S100B receptor and PI3K decreases C. difficile toxin-induced IL-6 upregulation in EGCs. (A) RAGE gene expression (mean ± s.e.m) by qPCR in enteroglial cells (EGC/PK060399) challenged with TcdA and TcdB (n = 4). (B) Analysis of IL-6 gene expression (mean ± s.e.m) by qPCR in enteroglial cells (EGC/PK060399) challenged with TcdA and TcdB for 18 h in the presence or absence of 30 μM FPSZM1 (a RAGE antagonist) which was added 1 h prior to C. difficile toxin challenge. (C) Representative photomicrographs of NFκBp65 (green) immunostaining and DAPI (blue) nuclear staining in enteroglial cells (EGC/PK060399) exposed to TcdA and TcdB after 18 h of incubation. (D) Percentages of cells (mean ± s.e.m) with positive nuclear NFκBp65 staining under different experimental conditions at 18 h of incubation with TcdA and TcdB. (E) Western blot (WB) bands of NFκB p65 and PCNA in nuclear extract fraction of enteroglial cells (EGC/PK060399) exposed to TcdA and TcdB at 18 h of incubation. (F) Analysis of IL-6 gene expression (mean ± s.e.m) by qPCR in enteroglial cells (EGC/PK060399) challenged with TcdA and TcdB for 18 h in the presence or absence of 10 μM LY294002 (a PI3K inhibitor) which was added 1 h prior to C. difficile toxin challenge. (A–F) Cells receiving only supplemented DMEM was applied as a control. (B, F) Experiments were performed with the same negative (Control) and positive controls (TcdA and TcdB). **p < 0.0001. ANOVA followed by (B, D, F) Turkey test was used. *p < 0.01.

Article Snippet: The sections were then incubated with an S100B antibody (NBP2-54426, Novus Biologicals, 1:1,000) overnight.

Techniques: Gene Expression, Immunostaining, Staining, Incubation, Western Blot, Control, Negative Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: S100B Inhibition Attenuates Intestinal Damage and Diarrhea Severity During Clostridioides difficile Infection by Modulating Inflammatory Response

doi: 10.3389/fcimb.2021.739874

Figure Lengend Snippet: Schematic diagram of the hypothetical role of S100B during C. difficile infection. C. difficile releases TcdA and TcdB which in turn promote epithelial damage and release of S100B by colonic EGCs. S100B stimulates the synthesis and secretion of inflammatory mediators (IL-1β, TNF-α, IL-18, IL-6, GMCSF, IL-17, IL-23, IL-2) promoting recruitment of immune cells, such as neutrophils, macrophages, and T cells, resulting in amplification of the C. difficile toxin-induced colonic damage. TcdA and TcdB induce IL-6 expression via S100B/RAGE/PI3K/NFκB. S100B also impairs epithelial integrity during CDI by decreasing IL-22 production, thereby hampering repair of the epithelium. Inhibition of S100B by pentamidine (PEN) blocks these events mediated by S100B during CDI.

Article Snippet: The sections were then incubated with an S100B antibody (NBP2-54426, Novus Biologicals, 1:1,000) overnight.

Techniques: Infection, Amplification, Expressing, Inhibition

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