Journal: Acta neuropathologica communications

Article Title: Polarizing receptor activation dissociates fibroblast growth factor 2 mediated inhibition of myelination from its neuroprotective potential.

doi: 10.1186/s40478-019-0864-6

Figure Lengend Snippet: Fig. 4 FGFR1 supports induction of pro-myelinating and neuroprotective factors. Mouse myelinating cultures were treated with 100 ng/ml FGF2 or F2 V2 at DIV18 for 3 days and protein levels in the cell culture supernatant were analyzed by ELISA for CXCL1 (a) and TIMP1 (b) or by Proteom Profiler (c, Mouse Cytokine Array Panel a). a, b FGF2 treatment results in a significant increase in protein levels for both CXCL1 and MMP3 (p < 0.05), whereas F2 V2 only significantly increased the levels of CXCL1. Data are presented as means + SEM from 6 independent experiments; * p < 0.05. c TIMP1 levels are increased in both FGF2 and F2 V2 treated cultures; data are presented as technical duplicates from 2 independent experiments, pixel densities are represented as arbitrary units. d-f A monolayer of neurosphere-derived mouse astrocytes was treated with 100 ng/ml recombinant FGF2 for 3 days and concentrations of CXCL1 (d), MMP3 (e) or TIMP1 (f) were determined by ELISA (d, e) or Proteom Profiler (f, Mouse Cytokine Array Panel a). d, e Data are presented as means ± SEM from 4 independent experiments; * p < 0.05; ** p < 0.01 (paired t-test), f Data are presented as technical duplicates from 2 independent experiments, arbitrary units represent pixel densities. g FGFR1 expression co- localizes with GFAP+ astrocytes in active multiple sclerosis. h, i Myelinating CNS cultures were treated in the absence or presence of 100 ng/ml FGF2 for 10 days from DIV 18 onwards and either directly fixed and stained for MOG / SMI31 after a total of 28 days (28 DIV) or after 38 days in vitro (38 DIV); shown are mean + SEM from at least 4 independent experiments and p-values for paired t-test; * p < 0.05. j Representative confocal immunofluorescence images (63x magnification, Maximum Intensity Projection) of 38 DIV culture demonstrating withdrawal of FGF2 (18 DIV – 28 DIV) is followed by rapid remyelination resulting in the formation of myelinated internodes that are indistinguishable from those in untreated cultures. Scale bars represent 10 μm

Article Snippet: Protein concentrations of CXCL1 and MMP3 in the supernatants were measured using mouse CXCL1 or MMP3 Quantikine ELISA kits (both R&D Systems) according to the manufacturer’s instructions.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Derivative Assay, Recombinant, Expressing, Staining, In Vitro, Immunofluorescence

Journal: PLoS ONE

Article Title: Stromal Cells Positively and Negatively Modulate the Growth of Cancer Cells: Stimulation via the PGE2-TNFα-IL-6 Pathway and Inhibition via Secreted GAPDH-E-Cadherin Interaction

doi: 10.1371/journal.pone.0119415

Figure Lengend Snippet: ( A ) MKN-7 and MKN-74 cells were cultured alone (Mono) or co-cultured with Hs738 (Cocul) for 3 days in the presence of MEK inhibitor I or U0126. Hs738 cells were cultured with the inhibitors for 2 days and CM was prepared. Both gastric cancer lines were cultured in the CM for 3 days. Cell growth was determined by measuring GFP fluorescence intensity. The values are means ± s.d. (n = 3). Cell growth is expressed as a percentage of the value without test compounds in each culture condition. ( B ) Hs738 cell extracts were incubated with b-MEK inh-pretreated streptavidin resin in the presence or absence of MEK inhibitor I (M) or U0126 (U) and the bound proteins were analyzed by Western blotting. L, 1/50 of loaded cell extracts. ( C ) Hs738 cells were cultured with MEK inhibitor I for 2 days and the cell lysates were analyzed by Western blotting. ( D ) Hs738 cells were cultured with MEK inhibitor I for 2 days and the concentrations of IL-6 and CXCL1 in the CM were determined. The values are means ± s.d. (n = 3). ( E ) Hs738 cells were treated with siRNA specific for RPL-18A (siRPL) or negative control (siCont) for 2 days and then re-inoculated followed by further culture for 2 days. The cell lysates were analyzed by Western blotting and the amounts of IL-6 in the CM were determined. The values are means ± s.d. (n = 3).

Article Snippet: Anti-human IL-6 neutralizing antibody (MAB206), recombinant human IL-6 (206-IL), and recombinant human CXCL1 (275-CR/CF) were purchased from R&D Systems.

Techniques: Cell Culture, Fluorescence, Incubation, Western Blot, Negative Control

Journal: PLoS ONE

Article Title: Stromal Cells Positively and Negatively Modulate the Growth of Cancer Cells: Stimulation via the PGE2-TNFα-IL-6 Pathway and Inhibition via Secreted GAPDH-E-Cadherin Interaction

doi: 10.1371/journal.pone.0119415

Figure Lengend Snippet: ( A ) MKN-7 and MKN-74 cells were cultured for 15 min with or without 50 μg/mL anti-IL-6 neutralizing antibody in Hs738 CM prepared by 2 days of culture. The activation of STAT3 was analyzed by Western blotting. ( B ) MKN-7 and MKN-74 cells were cultured for 15 min with IL-6. The activation of STAT3 was analyzed by Western blotting. ( C ) MKN-7 and MKN-74 cells were cultured with IL-6 or CXCL1 for 3 days. Cell growth was determined by measuring GFP fluorescence intensity. The values are means ± s.d. (n = 3). ( D ) MKN-7 and MKN-74 cells were cultured alone (Mono) or co-cultured with Hs738 cells (Cocul) with the indicated antibodies for 3 days. Cell growth was determined by measuring GFP fluorescence intensity. The values are means ± s.d. (n = 3). Cell growth is expressed as a percentage of the value without antibodies in each culture condition.

Article Snippet: Anti-human IL-6 neutralizing antibody (MAB206), recombinant human IL-6 (206-IL), and recombinant human CXCL1 (275-CR/CF) were purchased from R&D Systems.

Techniques: Cell Culture, Activation Assay, Western Blot, Fluorescence