Journal: Cell reports

Article Title: Suppressing Aneuploidy-Associated Phenotypes Improves the Fitness of Trisomy 21 Cells

doi: 10.1016/j.celrep.2019.10.059

Figure Lengend Snippet: (A) Representative images of RPE-1 nuclei mock treated and treated with myriocin for 24 h. Immunofluorescence for lamin B1 is red, and the nucleus is blue stained with Hoechst 33342. (B) Percentage of nuclear abnormalities of RPE-1 cells treated with myriocin (n = 100 cells). (C) Representative images of RPE-1 nuclei upon knockdown of SPTLC1 or SPTLC2 after 72 h. (D) Percentage of nuclear abnormalities of RPE-1 cells upon knockdown of SPTLC1 or SPTLC2 (n = 100 cells). (E) Western blot analysis of SPTLC1 and SPTLC2 protein levels in RPE-1 cells. (F) Representative images of RPE-1 nuclei treated with DHS, sphingosine (Sph), sphingosine-1-phosphate (S1P), or ceramide (Cer) for 24 h. (G) Percentage of RPE-1 cells showing abnormal nuclear morphology when treated with DHS, sphingosine (Sph), sphingosine-1-phosphate (S1P), or ceramide (n = 100 cells). (H) Representative images of primary human skin fibroblast (HSF) nuclei mock treated and treated with myriocin for 24 h. (I) Percentage of nuclear abnormalities of primary HSF treated with myriocin (n = 100 cells). (J) Representative images of primary HSF nuclei treated with DHS, sphingosine (Sph), sphingosine-1-phosphate (S1P), or ceramide for 24 h. (K) Percentage of primary HSF showing abnormal nuclear morphology when treated with DHS, sphingosine (Sph), sphingosine-1-phosphate (S1P), or ceramide (Cer) (n = 100 cells). All images are representative of three biological replicates. Scale bars (A, C, F, H, and J), 10 μm.

Article Snippet: Euploid human skin fibroblast (Control)_2 , Coriell Institute , GM08447; RRID:CVCL_7487.

Techniques: Immunofluorescence, Staining, Knockdown, Western Blot

Journal: Cell reports

Article Title: Suppressing Aneuploidy-Associated Phenotypes Improves the Fitness of Trisomy 21 Cells

doi: 10.1016/j.celrep.2019.10.059

Figure Lengend Snippet: (A) Representative images of primary human fibroblasts from 2 euploid donors and 2 patients with Down syndrome. Immunofluorescence for lamin B1 is red, and the nucleus is blue stained with Hoechst 33342. Scale bar, 2.5 μm. (B) Percentage of fibroblasts from euploid donors showing abnormal nuclear morphology when treated with fumonisin B 1 or SKi-II (n = 100). (C) Percentage of fibroblasts from 2 patients with Down syndrome showing abnormal nuclear morphology treated with fumonisin B 1 , SKi-II, or ceramide (Cer) (n = 100). (D) Representative images of trisomy 21 nuclei treated with fumonisin B 1 , SKi-II, or ceramide. Scale bar, 10 μm. (E) Representative images of primary human fibroblasts from 2 euploid donors and 2 patients with Patau syndrome(trisomy 13) or Edward syndrome (trisomy 18). Immunofluorescence for lamin B1 is red, and the nucleus is blue stained with Hoechst 33342. Scale bar, 2.5 μm. (F) Percentage of fibroblasts from 2 patients with Patau syndrome showing abnormal nuclear morphology treated with fumonisin B1, SKi-II, orceramide (n = 100). (G) Percentage of fibroblasts from 2 patients with Edward syndrome showing abnormal nuclear morphology treated with fumonisin B1, Ski-II, or ceramide (n = 100).

Article Snippet: Euploid human skin fibroblast (Control)_2 , Coriell Institute , GM08447; RRID:CVCL_7487.

Techniques: Immunofluorescence, Staining

Journal: Cell reports

Article Title: Suppressing Aneuploidy-Associated Phenotypes Improves the Fitness of Trisomy 21 Cells

doi: 10.1016/j.celrep.2019.10.059

Figure Lengend Snippet: (A) Representative images of primary human fibroblasts from euploid donors and patients with Patau syndrome (trisomy 13), Edward syndrome (trisomy 18), or Down syndrome (trisomy 21). Immunofluorescence is shown for lamin A/C, H3K9triMe, and Hoechst 33342. In the merge images, lamin A/C is green, H3K9triMe is red, and DNA is blue. Scale bar, 5 μm. (B) Western blot analysis of H3K9triMe in human fibroblasts. (C) Quantification of the number of 53BP1 foci in human fibroblasts mock treated or in the presence of fumonisin B 1 (n = 100). (D) Representative images of primary human fibroblasts from euploid donors and patients with Down syndrome. Immunofluorescence is shown for 53BP1 (red) and Hoechst 33342 (blue). There is no correlation between abnormal nuclear morphology and number of 53BP1 foci. Scale bar, 5 μm. (E) Growth curves of primary human fibroblasts from 2 euploid donors and 2 patients with Patau (trisomy13), Edward (trisomy18), or Down syndrome(trisomy 21) with increasing concentrations of fumonisin B 1 . (F) Representative images of primary human astrocytes from 4 euploid donors and 4 donors with Down syndrome. Immunofluorescence for the nucleus is blue stained with Hoechst 33342. Scale bar, 5 μm. (G) Percentage of astrocytes from euploid and trisomy 21 donors showing abnormal nuclear morphology (n > 200).

Article Snippet: Euploid human skin fibroblast (Control)_2 , Coriell Institute , GM08447; RRID:CVCL_7487.

Techniques: Immunofluorescence, Western Blot, Staining

Journal: Cell reports

Article Title: Suppressing Aneuploidy-Associated Phenotypes Improves the Fitness of Trisomy 21 Cells

doi: 10.1016/j.celrep.2019.10.059

Figure Lengend Snippet:

Article Snippet: Euploid human skin fibroblast (Control)_2 , Coriell Institute , GM08447; RRID:CVCL_7487.

Techniques: Recombinant, Membrane, Protease Inhibitor, Control, Software

Journal: Journal of Cellular and Molecular Medicine

Article Title: Oncostatin M decreases interleukin-1 β secretion by human synovial fibroblasts and attenuates an acute inflammatory reaction in vivo

doi: 10.1111/j.1582-4934.2011.01412.x

Figure Lengend Snippet: Impact of OSM on gene expression (A, B) and protein secretion (C, D) in stimulated HSFs. HSFs were stimulated for 24 hrs with a combination of LPS (100 ng/ml) and GM-CSF (1.4 nM). Human recombinant OSM (10 ng/ml) was added with or without 10 μg/ml of neutralizing anti-OSM, or isotype-matched non-specific antibody. Levels of IL-1β (A), IL-6, CXCL8 and CCL2 mRNA (B) were measured by real-time PCR. Results (mean ± S.E.M., n ∇ 6) are expressed as multiples relative to stimulated cells (dotted line). Protein levels of IL-1β (C), IL-6, CCL2 and CXCL8 (D) in supernatants were determined by ELISA (mean ± S.E.M., n ∇ 6). *Significantly different ( P < 0.05) from stimulated cells. # Significantly different from the stimulated cell in the presence of OSM. Control: unstimulated cells.

Article Snippet: Normal human primary synovial fibroblasts (HSFs) were purchased from Asterand (Detroit, MI, USA).

Techniques: Gene Expression, Recombinant, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Oncostatin M decreases interleukin-1 β secretion by human synovial fibroblasts and attenuates an acute inflammatory reaction in vivo

doi: 10.1111/j.1582-4934.2011.01412.x

Figure Lengend Snippet: Impact of PMN-derived OSM on gene expression and protein secretion in stimulated HSFs. PMNs were stimulated with a combination of LPS (100 ng/ml) and GM-CSF (1.4 nM) for 4 hrs. (A) OSM concentrations in supernatants of resting or LPS/GM-CSF-stimulated PMNs were measured by ELISA (mean ± S.E.M., n ∇ 3). (B) HSFs were incubated for 24 hrs with supernatants of resting or stimulated PMNs with or without anti-OSM neutralizing antibody (10 μg/ml). Expression levels are presented relative to the levels for cells stimulated in the absence of anti-OSM (as reciprocals in the case of decreases), as measured using real-time PCR (mean ± S.E.M., n ∇ 4). (C) IL-1β, IL-6, CCL2 and CXCL8 concentrations were quantified in cell-free supernatants by ELISA. Results are expressed as mean ± S.E.M. ( n ∇ 4). *Significantly different from SRP. # Significantly different ( P < 0.05) from SSP.

Article Snippet: Normal human primary synovial fibroblasts (HSFs) were purchased from Asterand (Detroit, MI, USA).

Techniques: Derivative Assay, Gene Expression, Enzyme-linked Immunosorbent Assay, Incubation, Expressing, Real-time Polymerase Chain Reaction

Journal: Journal of Cellular and Molecular Medicine

Article Title: Oncostatin M decreases interleukin-1 β secretion by human synovial fibroblasts and attenuates an acute inflammatory reaction in vivo

doi: 10.1111/j.1582-4934.2011.01412.x

Figure Lengend Snippet: Response of HSF mRNA levels to LIF and IL-6. HSFs were stimulated with a combination of LPS and GM-CSF for 24 hrs in the absence or presence of OSM, LIF or IL-6 (10 ng/ml each). Levels of IL-1β, IL-6, CXCL8 and CCL2 mRNA were quantified by real-time PCR. Results (mean ± S.E.M., n ∇ 3) are expressed as multiples relative to stimulated cells (dotted line). * P < 0.05, ** P < 0.01 from the stimulated condition.

Article Snippet: Normal human primary synovial fibroblasts (HSFs) were purchased from Asterand (Detroit, MI, USA).

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of Cellular and Molecular Medicine

Article Title: Oncostatin M decreases interleukin-1 β secretion by human synovial fibroblasts and attenuates an acute inflammatory reaction in vivo

doi: 10.1111/j.1582-4934.2011.01412.x

Figure Lengend Snippet: Signalling pathways mediating the OSM-effects on the IL-1β, CXCL8, IL-6 and CCL2 expression in stimulated HSFs. (A) HSFs were stimulated or not with a combination of LPS and GM-CSF for 10 min. in the absence or presence of OSM (10 ng/ml) and its neutralizing antibody (10 μg/ml), as indicated, prior to lysis and subsequent immunoblot analysis using primary antibodies raised against phospho-ERK1/2, phospho-STAT-1 (Tyr 701) and total STAT-1 (as a loading control). Immunoblots shown are representative of three independent experiments; bands were quantified by densitometric analysis and results presented as mean ± S.E.M. When indicated, cells were pre-incubated for 15 min. with the MEK inhibitor U0126 (10 μM) or the Jak inhibitor (0.5 μM) before stimulation in the presence of OSM. * P < 0.05 from the control condition (fixed at 1). (B) Otherwise identical conditions were used to study the consequences of a 24-hr stimulation in the presence of OSM and the metabolic inhibitors, on the mRNA expression of IL-1β, CXCL8, IL-6 and CCL2 by real-time PCR. Results (mean ± S.E.M., n ∇ 4) are expressed as multiples relative to stimulated cells (dotted line). * P < 0.05 from the stimulated condition, # P < 0.05 from the s. + OSM condition. (C) Schematic representation of the OSMRβ signalling in LPS and GM-CSF stimulated HSFs. OSM receptor engagement induces activation/phosphorylation of STAT-1 and also increases ERK1/2 phosphorylation. This leads to a STAT-1-dependent negative regulation of the IL-1β and CXCL8 genes transcription and to the up-regulation of the IL-6 and CCL2 genes.

Article Snippet: Normal human primary synovial fibroblasts (HSFs) were purchased from Asterand (Detroit, MI, USA).

Techniques: Expressing, Lysis, Western Blot, Control, Incubation, Real-time Polymerase Chain Reaction, Activation Assay, Phospho-proteomics

Journal: Nanomaterials

Article Title: Development and Biocompatibility Evaluation of Photocatalytic TiO 2 /Reduced Graphene Oxide-Based Nanoparticles Designed for Self-Cleaning Purposes

doi: 10.3390/nano7090279

Figure Lengend Snippet: Representative images of phase contrast microscopy revealing cellular morphology of CCD 1070Sk ( a ) and MRC-5 ( b ) fibroblasts exposed to 100 and 200 μg/mL of two TiO 2 /RGO nanocomposites samples (A and B) and commercially available TiO 2 NPs (P25) for 24 and 48 h. Scale bar: 100 μm.

Article Snippet: Normal human lung (MRC-5 cell line, ATCC Cat. No. CCL-171) and skin (CCD 1070Sk cell line, ATCC Cat. No. CRL-2091) fibroblasts were cultured in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with an addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C, in a 5% CO 2 humidified atmosphere.

Techniques: Microscopy

Journal: Nanomaterials

Article Title: Development and Biocompatibility Evaluation of Photocatalytic TiO 2 /Reduced Graphene Oxide-Based Nanoparticles Designed for Self-Cleaning Purposes

doi: 10.3390/nano7090279

Figure Lengend Snippet: Lysosomes profile in skin (CCD 1070Sk) and lung (MRC-5) fibroblasts after 48 h exposure to different concentrations (100 and 200 μg/mL) of the two TiO 2 /RGO nanocomposites samples (A and B) and commercially available TiO 2 (P25). Lysosomes (green) staining was done with LysoTracker Green DND-26 and nuclei (blue) were counterstained with Hoechst 33342. Scale bar: 100 μm.

Article Snippet: Normal human lung (MRC-5 cell line, ATCC Cat. No. CCL-171) and skin (CCD 1070Sk cell line, ATCC Cat. No. CRL-2091) fibroblasts were cultured in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with an addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C, in a 5% CO 2 humidified atmosphere.

Techniques: Staining