Journal: bioRxiv

Article Title: IL-1-conferred gene expression pattern in ERα + BCa and AR + PCa cells is intrinsic to ERα − BCa and AR − PCa cells and promotes cell survival

doi: 10.1101/773978

Figure Lengend Snippet: RT-qPCR was performed for select genes for MCF7 and LNCaP cell lines treated with vehicle control or 25 ng/ml IL-1α or IL-1β for 5 days. MDA-MB-231, BT549, PC3 and DU145 cell lines were treated with vehicle control only. (A) RT-qPCR shows that CCL20 , CD68 , IL8 , p62 , SOX9 and Zeb1 are induced by IL-1 in MCF7 and/or LNCaP cell lines and are basally high in MDA-MB-231, BT549, PC3 and/or DU145 cell lines. (B) RT-qPCR shows that CXCR7 and MMP16 , but not CDK2 or PLK1 , are downregulated by IL-1 in MCF7 and/or LNCaP cell lines and are basally high in MDA-MB-231, BT549, PC3 and/or DU145 cell lines. N = 3 biological replicates; error bars, +/−STDEV; p-value, * ≤ 0.05, ** ≤ 0.005, ***≤ 0.0005. mRNA fold change is normalized to MCF7 vehicle control for the BCa cell lines and to LNCaP vehicle control for the PCa cell lines.

Article Snippet: Human recombinant IL-1α (GoldBio, St Louis MO; 1110-01A-100) and IL-1β (GoldBio, St Louis MO; 1110-01B-100) were resuspended in 0.1% bovine serum albumin (BSA, Thermo Fisher Scientific; BP 1600-1) in 1X phosphate buffered saline (PBS).

Techniques: Quantitative RT-PCR

Journal: American journal of physiology. Endocrinology and metabolism

Article Title: Myostatin propeptide mutation of the hypermuscular Compact mice decreases the formation of myostatin and improves insulin sensitivity.

doi: 10.1152/ajpendo.00216.2016

Figure Lengend Snippet: Fig. 3. Myostatin level in skeletal muscle of Compact, congenic wild-type, and BALB/c mice. M. gastrocnemius protein extracts were subjected to SDS-PAGE and blotted with anti-myostatin or anti-propeptide antibody. Represen- tative images are shown. Note the presence of mature myostatin dimer and myostatin propeptide in Compact sam- ples. Mouse recombinant myostatin was used as a positive control, and muscle homogenates of myostatin knockout (KO) mice served as a negative control. Differences in glycosylation may cause altered electrophoretic mobility. Bar diagrams show the quantification of the results. Data are reported as means SE; n 5 Compact, 5 congenic wild-type, and 6 BALB/c mice. *P 0.05; **P 0.01; ***P 0.001.

Article Snippet: Mouse recombinant myostatin (788-G8; R&D Systems) was used as a positive control.

Techniques: SDS Page, Recombinant, Positive Control, Knock-Out, Negative Control, Glycoproteomics

Journal: American journal of physiology. Endocrinology and metabolism

Article Title: Myostatin propeptide mutation of the hypermuscular Compact mice decreases the formation of myostatin and improves insulin sensitivity.

doi: 10.1152/ajpendo.00216.2016

Figure Lengend Snippet: Fig. 5. Glucose tolerance and insulin sensitivity are improved by Compact myostatin mutation and reduced in congenic wild-type mice. Intraperitoneal (ip) glucose tolerance (A and B) and insulin sensitivity tests (C and D) of 3- to 4-mo-old (A and C) and 10-mo-old animals (B and D). Area under the curve (AUC) values are presented in bar diagrams. Data are reported as means SE. *P 0.05 and **P 0.01; n 3 Compact, 7 congenic wild-type, and 3 BALB/c mice (A), n 7 Compact, 3 congenic wild-type, and 6 BALB/c mice (B), n 3 Compact, 3 congenic wild-type, and 4 BALB/c mice (C), and n 6 Compact, 4 congenic wild-type, and 4 BALB/c mice (D).

Article Snippet: Mouse recombinant myostatin (788-G8; R&D Systems) was used as a positive control.

Techniques: Mutagenesis

Journal: Frontiers in Cell and Developmental Biology

Article Title: Meclozine Attenuates the MARK Pathway in Mammalian Chondrocytes and Ameliorates FGF2-Induced Bone Hyperossification in Larval Zebrafish

doi: 10.3389/fcell.2021.694018

Figure Lengend Snippet: Meclozine attenuates the MAPK pathway of FGF2-treated tibiae in the organ culture system. (A) Upper panels: Representative images of E16.5 tibiae of wild-type mice after 4-day culture. Scale bares indicate 1 mm. Lower panels: Absolute bone length after 4-day treatment. Dots indicate the length. Lines are drawn between dots of the same individual. Statistical significance was analyzed by paired Student’s t -test. (B) Enrichment plots of two MAPK signaling-associated gene sets identified by GSEA between FGF2- and FGF2+, and FGF2+ and FGF2+ meclozine in the articular cartilage of ex vivo cultured tibiae. (C) Heatmap depicting the expression of the genes in REACTOME_MAPK_FAMILY_SIGNALING_CASCADES signature and ST_P38_MAPK_PATHWAY signature between FGF2- and FGF2+, and FGF2+ and FGF2+ meclozine in the articular cartilage of ex vivo cultured tibiae. (D) Enrichment plots of BMP signaling-associated gene set identified by GSEA between FGF2- and FGF2+, and FGF2+ and FGF2+ meclozine in the articular cartilage of ex vivo cultured tibiae. (E) Heatmap depicting the expression of the Ihh , Bmp2 , Bmp4 , and Bmp7 between FGF2- and FGF2+, and FGF2+ and FGF2+ meclozine in the articular cartilage of ex vivo cultured tibiae. MAPK, mitogen-activated protein kinase; GSEA, Gene set enrichment analysis.

Article Snippet: FGF2 (3339-FB-025, R and D Systems) was administered in the presence or absence of 20 μM meclozine (155341, MP Biomedicals) for 4 days.

Techniques: Organ Culture, Ex Vivo, Cell Culture, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Meclozine Attenuates the MARK Pathway in Mammalian Chondrocytes and Ameliorates FGF2-Induced Bone Hyperossification in Larval Zebrafish

doi: 10.3389/fcell.2021.694018

Figure Lengend Snippet: Treatment protocol of FGF2 and meclozine are determined for evaluating vertebral ossification in larval zebrafish. (A) Treatment regimen of FGF2 for larval zebrafish. (B) Quantification of ossified vertebrae after FGF2 treatment. Dots indicate the number of ossified vertebrae of each sample, and bars indicate means. Statistical significance was analyzed by one-way ANOVA with post-hoc Tukey HSD. (C) Representative images of larval zebrafish from the lateral view at seven dpf stained with Alizarin red after 30 ng/mL FGF2 treatment from eight hpf to seven dpf. Arrows: ossified vertebrae. Scale bar indicates 500 µm. (D) Survival curve of larval zebrafish treated with each dose of meclozine from eight hpf to seven dpf.

Article Snippet: FGF2 (3339-FB-025, R and D Systems) was administered in the presence or absence of 20 μM meclozine (155341, MP Biomedicals) for 4 days.

Techniques: Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Meclozine Attenuates the MARK Pathway in Mammalian Chondrocytes and Ameliorates FGF2-Induced Bone Hyperossification in Larval Zebrafish

doi: 10.3389/fcell.2021.694018

Figure Lengend Snippet: Meclozine attenuates spinal and craniofacial bone ossification in FGF2-treated larval zebrafish. (A) Representative images of larval zebrafish from the lateral view at seven dpf stained with Alizarin red after 30 ng/mL FGF2 treatment, with or without 1 µM meclozine, from eight hpf to seven dpf. Arrows: ossified vertebrae. Scale bar indicates 500 µm. (B) Quantification of ossified vertebrae after FGF2 treatment, with or without meclozine. Dots indicate the number of ossified vertebrae of each sample, and bars indicate means. Statistical significance was analyzed by one-way ANOVA with post-hoc Tukey HSD. hpf, hours post-fertilization; dpf, days post-fertilization. (C) Representative craniofacial bone elements of larval zebrafish from anteroposterior view at seven dpf stained with Alizarin red after FGF2 treatment, with or without meclozine. Scale bar indicates 500 µm. (D) Quantification of the number of each ossified craniofacial bone element, including ceratohyal (ch), hyomandibular (hm), branchiostegal ray (br), dentary (d), entopterygoid (en), maxilla (m), and opercle (o), after FGF2 treatment with or without meclozine. Data values are presented as means and standard deviation (SD). Statistical significance was analyzed by one-way ANOVA with post-hoc Tukey HSD.

Article Snippet: FGF2 (3339-FB-025, R and D Systems) was administered in the presence or absence of 20 μM meclozine (155341, MP Biomedicals) for 4 days.

Techniques: Staining, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Meclozine Attenuates the MARK Pathway in Mammalian Chondrocytes and Ameliorates FGF2-Induced Bone Hyperossification in Larval Zebrafish

doi: 10.3389/fcell.2021.694018

Figure Lengend Snippet: Meclozine ameliorates FGF2-induced hyper ossification in larval zebrafish. (A) Representative craniofacial cartilage elements of larval zebrafish from ventral view, three-dimensional (3D) view, and single layer at seven dpf in Tg (col2a1a:EGFP) after FGF2 treatment, with or without meclozine. Scale bar indicates 100 µm. (B) Quantification of area of craniofacial cartilage, including ceratohyal (ch), hyosymplectic (h), and palatoquadrate (pq) after FGF2 treatment with or without meclozine. (C) Representative craniofacial cartilage and bone elements of larval zebrafish from ventral view, 3D view, and single layer at seven dpf in Tg (col2a1a:EGFP) stained with Alizarine red after FGF2 treatment, with or without meclozine. Scale bar indicates 100 µm. (D) Quantification of relative ossification area, including ceratohyal (ch), hyomandibular (hm), and quadrate (q) after FGF2 treatment with or without meclozine. Relative ossification area was calculated by dividing each red signal area by each green area. Data values are presented as means and standard deviation (SD). Statistical significance was analyzed by one-way ANOVA with post-hoc Tukey HSD.

Article Snippet: FGF2 (3339-FB-025, R and D Systems) was administered in the presence or absence of 20 μM meclozine (155341, MP Biomedicals) for 4 days.

Techniques: Staining, Standard Deviation

Journal: CNS Neuroscience & Therapeutics

Article Title: A nerve conduit filled with Wnt5a‐loaded fibrin hydrogels promotes peripheral nerve regeneration

doi: 10.1111/cns.13752

Figure Lengend Snippet: Characterization of the Wnt5a‐loaded fibrin hydrogel. (A–B) Gross view of the saline solution (left) and fibrin hydrogel (right) at upright and upside‐down position. (C) A representative photograph of the nerve conduit filled with the Wnt5a‐loaded fibrin hydrogel. (D) Representative SEM images. (E) The absorbance of Wnt5a when the mass ratio (w/w) of Wnt5a and fibrin hydrogel was 1.2, 1.4, 1.6, and 1.8, respectively. (F) The loading capacity (w/w%) of Wnt5a‐loaded fibrin hydrogel in each group. (G) The sustained‐release profile of Wnt5a‐loaded fibrin hydrogel

Article Snippet: We mixed 100 UI/mL thrombin (T4648, Sigma‐Aldrich) and 0.8% fibrinogen (f8630, Sigma‐Aldrich, T4648, Sigma‐Aldrich) solutions at a ratio of 1:4 to form the fibrin hydrogel., We created a Wnt5a‐loaded fibrin hydrogel by mixing the Wnt5a solution (7815‐NG, R&D Systems) in thrombin and fibrinogen solutions, respectively, before the fibrin hydrogel was synthesized.

Techniques: Saline

Journal: CNS Neuroscience & Therapeutics

Article Title: A nerve conduit filled with Wnt5a‐loaded fibrin hydrogels promotes peripheral nerve regeneration

doi: 10.1111/cns.13752

Figure Lengend Snippet: Wnt5a regulates SC proliferation and secretion. (A–C) qRT‐PCR results showed relative expression levels of VEGF, NGF, and CNTF mRNA. (D–F) ELISA results indicated the concentrations of VEGF, NGF, and CNTF. (G) CCK‐8 results showed the effect of Wnt5a on SC proliferation after incubation for 24 h and 48 h.* p < 0.05, ** p < 0.01, *** p < 0.001. Data are expressed as the mean ±SD. SCs, Schwann cells; VEGF, vascular endothelial growth factor; NGF, nerve growth factor; CNTF, cholinergic neurotrophic factor. qRT‐PCR, quantitative real‐time polymerase chain reaction

Article Snippet: We mixed 100 UI/mL thrombin (T4648, Sigma‐Aldrich) and 0.8% fibrinogen (f8630, Sigma‐Aldrich, T4648, Sigma‐Aldrich) solutions at a ratio of 1:4 to form the fibrin hydrogel., We created a Wnt5a‐loaded fibrin hydrogel by mixing the Wnt5a solution (7815‐NG, R&D Systems) in thrombin and fibrinogen solutions, respectively, before the fibrin hydrogel was synthesized.

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Incubation, Real-time Polymerase Chain Reaction