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: Oncogene

Article Title: Transduction of the SkBr3 breast carcinoma cell line with the HOXB7 gene induces bFGF expression, increases cell proliferation and reduces growth factor dependence.

doi: 10.1038/sj.onc.1201875

Figure Lengend Snippet: Figure 1 Expression of HOXB7 and bFGF genes in A375 melanoma, SkBr3 mammary carcinoma and HOXB7-transduced SkBr3. (a) RNase protection analysis. Riboprobes (R) and protected fragments (P) are shown on the right, and molecular size (base pairs) markers (MWM) (pGEM4Z HpaII) on the left. b-actin expression is shown as internal control. (b) Northern blot analysis. b2 microglobulin expression is shown as internal control. (c) Western blot analysis of bFGF produced by A375 melanoma, SkBr3 mammary carcinoma and HOXB7-transduced SkBr3 cells

Article Snippet: Intracellular and secreted bFGF were quanti®ed by a human bFGF ELISA from R&D systems (Minneapolis, MN).

Techniques: Expressing, Control, Northern Blot, Western Blot, Produced

Journal: Oncogene

Article Title: Transduction of the SkBr3 breast carcinoma cell line with the HOXB7 gene induces bFGF expression, increases cell proliferation and reduces growth factor dependence.

doi: 10.1038/sj.onc.1201875

Figure Lengend Snippet: Figure 3 (a) Role of bFGF for SkBr3/HOXB7 cell growth in low serum. Growth kinetics of parental and HOXB7-transduced SkBr3 cells maintained in 10% or 1% FCS were compared. Cell growth was monitored over the indicated time intervals by methylene blue inclusion, as indicated in Materials and methods. (b) Eect of exogenous rbFGF on SkBr3 cell growth. Growth curves of cells maintained in 1% FCS plus dierent concentra- tions of rbFGF evaluated by methylene blue inclusion

Article Snippet: Intracellular and secreted bFGF were quanti®ed by a human bFGF ELISA from R&D systems (Minneapolis, MN).

Techniques:

Journal: Oncogene

Article Title: Transduction of the SkBr3 breast carcinoma cell line with the HOXB7 gene induces bFGF expression, increases cell proliferation and reduces growth factor dependence.

doi: 10.1038/sj.onc.1201875

Figure Lengend Snippet: Figure 4 Inhibition of SkBr3/HOXB7 cell proliferation upon treatment with 30 mM of bFGF antisense (a) or of sense (s) oligomers. Evidence of bFGF intracrine loop operating in cells kept in low serum. Mean+s.d. of three separate experiments is shown

Article Snippet: Intracellular and secreted bFGF were quanti®ed by a human bFGF ELISA from R&D systems (Minneapolis, MN).

Techniques: Inhibition

Journal: The Journal of Biological Chemistry

Article Title: Reactivation of Mitogen-activated Protein Kinase (MAPK) Pathway by FGF Receptor 3 (FGFR3)/Ras Mediates Resistance to Vemurafenib in Human B-RAF V600E Mutant Melanoma *

doi: 10.1074/jbc.M112.377218

Figure Lengend Snippet: Enhanced FGFR3 activation in vemurafenib-resistant B-RAF V600E melanoma cells. A, phospho-RTK antibody array analysis. Cell lysates from A375, M14, A375-R1, and M14-R cell lines were incubated on RTK antibody array for 16 h and phosphorylation status was determined as described under “Experimental Procedures.” Each RTK antibody is spotted in duplicate. Supplemental Table S2 describes the list of RTKs and the layout of the antibody array. B, confirmation of phospho-FGFR3 levels by Western blot analysis. Protein levels of total and phosho-FGFR3 were assessed using immunoblotting. C, ELISA analysis of secreted FGF2 in the conditioned media obtained from A375, A375-R1, M14, and M14-R cells. ELISA was performed as described in “Experimental Procedures.”

Article Snippet: Cells were cultured for 48 h in growth medium described above, and the conditioned medium samples (cell free culture supernatant) were analyzed for concentrations of human FGF2 using human FGF2 Quantikine ELISA Kit (R&D Systems).

Techniques: Activation Assay, Ab Array, Incubation, Phospho-proteomics, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Materials & Design

Article Title: Multifunctional nanofiber-based dressings in coordination with adipose-derived stem cells for accelerated burn wound healing

doi: 10.1016/j.matdes.2025.113929

Figure Lengend Snippet: Fig. 4. (A) Detection of ADSCs proliferation by ADM coating at different times. Quantification of (B) EGF and (C) FGF2 in ADSCs paracrine products. *** P < 0.001 representing a significant difference as compared with ADM (3 min) and TLWDA.

Article Snippet: Human EGF enzyme-linked immunosorbent assay (ELISA) kit and human FGF2 ELISA kit were purchased from BOSTER (Wuhan, China).

Techniques: