Journal: Nature Communications

Article Title: Identification of recurrent FHL2-GLI2 oncogenic fusion in sclerosing stromal tumors of the ovary

doi: 10.1038/s41467-019-13806-x

Figure Lengend Snippet: a Differential gene expression analysis of human sclerosing stromal tumors (SSTs) subjected to RNA-sequencing ( n = 8, this study) and high-grade serous ovarian carcinomas ( n = 16; The Cancer Genome Atlas) and other sex cord-stromal tumors (SCSTs, n = 11; this study). Gene expression fold-change is color-coded according to the legend. Only genes significantly differentially expressed ( P < 0.05; two-tailed unpaired t -test) are shown. CPM, count per million. b Expression levels of Sonic Hedgehog (SHH) pathway genes in human SSTs ( n = 11) and other sex-cord stromal tumors ( n = 9) as defined using NanoString. Expression levels and SHH enrichment scores are color-coded according to the legends. *** P < 0.001, Wilcoxon rank test. Hierarchical clustering was performed using complete linkage and Euclidian distance. c Quantitative assessment of the Sonic Hedgehog pathway PTCH1 and GLI1 transcripts in immortalized mesenchymal stem cells (MSCs), HEK-293 and medulloblastoma (DAOY) cells and of PTCH1 and CCND1 transcripts in human basal cell carcinoma (BCC) cells stably expressing empty vector (control), wild-type FHL2 (FHL2), wild-type GLI2 (GLI2), truncated GLI2 (tGLI2), or FHL2-GLI2. Expression levels were normalized to GAPDH expression, and comparisons of mRNA expression levels were performed relative to control. d Representative western blot analysis of PTCH1 and GLI1 protein expression in MSC, HEK-293 and DAOY cells and of PTCH1 and CCND1 in BCC cells stably expressing control, FHL2, GLI2, tGLI2, or FHL2-GLI2. Tubulin was used as protein loading control. Quantification (below) of protein levels as compared to control. In c – d , data are representative of at least three independent experiments. Error bars, s.d. of mean; n.s., not significant; * P < 0.05, ** P < 0.01, *** P < 0.001; two-tailed unpaired t -test.

Article Snippet: BCC cells were cultured in Human Basal Cell Carcinoma Cell Line Complete Media (CELPROGEN).

Techniques: Expressing, RNA Sequencing Assay, Two Tailed Test, Stable Transfection, Plasmid Preparation, Western Blot

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Skeletal colonization by breast cancer cells is stimulated by an osteoblast and β2AR-dependent neo-angiogenic switch

doi: 10.1002/jbmr.3133

Figure Lengend Snippet: A) Representative 40× H&E images of hind limb bone sections from athymic nude mice treated with PBS, 3mg/kg Isoproterenol (ISO), or Chronic Immobilization Stress (CIS). Blood vessels can be recognized by their shape and staining of red blood cells (red). Bar: 100 μm. B) Quantification of vessel area (VsA) and vessel number normalized by tissue area (TA) in mice that received PBS (N=12) or ISO (N=7) (p=.009 and p=.037), as well as mice subjected to control (N=5) or CIS (N=6) (p=.296 and p=.004). C) Quantification of primary mouse BMEC tube length (*=p<.05, N=3). D) Quantification of HUVECs tube length (*p=<.05, ***=p<.001, N=6). E) Schematic of the metatarsal assay and representative 4× Images of metatarsals according to each treatment. After 2 weeks of culture, mouse explants were stained for the endothelial cell marker CD31. F) Quantification of the number of CD31+ parental vessels sprouting from the metatarsal bones (*P=<.05, ****P=<.0001, N≥3).

Article Snippet: Twenty-four well plates were coated with 200 μL of growth factor reduced matrigel (Corning #354230) for 45 mins. at 37°C before 30,000 HUVECs (Gibco #C0035C), or BMECs from wild-type C57BL/6 mice were plated and cultured in 500 μL of Basal Endothelial Cell Growth Media (ScienCell #1001).

Techniques: Staining, Control, Marker

Journal: Toxics

Article Title: Nonylphenol Polyethoxylates Enhance Adipose Deposition in Developmentally Exposed Zebrafish

doi: 10.3390/toxics10020099

Figure Lengend Snippet: Nonylphenol and polyethoxylates promote adipogenesis in human mesenchymal stem cell models. Zenbio and Lonza human bone marrow–derived mesenchymal stem cell models were differentiated as described in Methods and assessed for adipocyte differentiation (Nile Red staining of lipid accumulation) and cell proliferation (Hoechst staining) after 14/21 (respectively) days of differentiation while exposed to controls chemicals as well as nonylphenol and its ethoxylates. Percent normalized triglyceride accumulation per cell relative to maximal rosiglitazone response (normalized to DNA content) ( A , C ). increase (cell proliferation) or decrease (potential cytotoxicity) in DNA content relative to vehicle control ( B , D ). Zenbio–sourced cell data provided in ( A , B ), and Lonza–sourced cell data provided in ( C , D ). Data presented as mean ± SEM from three independent experiments. * indicates lowest concentration with significant increase in triglyceride over vehicle control or cell proliferation/cytotoxicity relative to vehicle control, p < 0.05, as per Kruskal–Wallis in GraphPad Prism 9. X–axis format is provided in log scale. Panel ( E ) provides a summary plot of maximal effects on triglyceride accumulation based on ethoxylate chain length across cell models, comparing results from panels ( A , C ) with previously published effects in 3T3–L1 cells (PMID: 29106673). NPEO = nonylphenol polyethoxylated (with varying average ethoxylate chain lengths).

Article Snippet: Cells were maintained as suggested by manufacturer in manufacturer-specific mesenchymal stem cell basal growth media for Zenbio (catalog #BMSC-1) and for Lonza (catalog #PT-3001).

Techniques: Derivative Assay, Staining, Control, Concentration Assay