Journal: Cell reports

Article Title: Variation of Human Neural Stem Cells Generating Organizer States In Vitro before Committing to Cortical Excitatory or Inhibitory Neuronal Fates

doi: 10.1016/j.celrep.2020.107599

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: 1 × 10 6 cells were plated over 10 cm culture plates (Falcon, 35–3003), previously coated with poly-L-ornithine (PLO) (Sigma, P3655) and fibronectin (FN) (R&D Systems, 1030FN), and incubated at 37°C, 5% O 2 and 5% CO 2 for 5 days in DMEM/F12 medium (Mediatech 16–405-CV) plus N2 supplement, containing 25 μg/ml bovine insulin (Sigma, I6634), 100 μg/mL apotransferrin (Sigma, T2036), 20 nM progesterone (Sigma, P8783), 100 mM putrescine (Sigma, P5780), 30 nM sodium selenite (Sigma, S5261), penicillin/streptomycin (Life Technology, 15140–122).

Techniques: Virus, Plasmid Preparation, Recombinant, Transfection, Antibody Labeling, In Vitro, Microarray, Gene Expression, Derivative Assay, Software

Journal: eLife

Article Title: Tumor stiffening reversion through collagen crosslinking inhibition improves T cell migration and anti-PD-1 treatment

doi: 10.7554/eLife.58688

Figure Lengend Snippet: Migration of activated PBT plated onto fresh tumor slices was analyzed in EGI-1 and MMTV-PyMT tumor model, whilst resident tumor-infiltrating T lymphocytes were analyzed in mPDAC and KPC tumor model. Illustrative images of T cell migration tracks in EGI-1, MMTV-PyMT, mPDAC, and KPC tumor models. Tumor stroma (fibronectin) in red, tumor cells (EpCAM in EGI-1, MMTV-PyMT, and KPC tumor models, CD44 in mPDAC tumor models), in blue and T cells (CD8 in mPDAC and KPC, Calcein in MMTV-PyMT, and EGI-1 tumor models) in green. Tracks are color-coded to illustrate track displacement. Scale bar = 100 µm. T cell migration speed, T cell displacement, and trajectory straightness in all tumor models. ***p - value>0.001, p-value>0.05, Student’s t-test. Results are shown as mean ± SD. Figure 5—source data 1. Source data file for .

Article Snippet: Live vibratome sections were stained with BV421 anti-mouse EpCAM (G8.8 clone; BD Biosciences), PerCP-e710 anti-mouse CD8a (53–6.7 clone, eBioscience), PE anti-podoplanin (8.1.1 clone; BioLegend), and anti-human/mouse fibronectin (HFN7.1 clone; Novus Biologicals).

Techniques: Migration

Journal: eLife

Article Title: Tumor stiffening reversion through collagen crosslinking inhibition improves T cell migration and anti-PD-1 treatment

doi: 10.7554/eLife.58688

Figure Lengend Snippet: ( A ) Representative images of Control and BAPN-treated KPC tumors. Tumor cells (EpCAM) in blue, tumor stroma (fibronectin) in red, and T cells (CD8) in green. Scale bar = 100 µm. ( B ) Quantification of CD8 + T cells per mm 2 in tumor islets and stroma regions in control and BAPN (*p<0.01, Student’s t-test, n = 2–3 mice/group). Figure 5—figure supplement 1—source data 1. Source data file for .

Article Snippet: Live vibratome sections were stained with BV421 anti-mouse EpCAM (G8.8 clone; BD Biosciences), PerCP-e710 anti-mouse CD8a (53–6.7 clone, eBioscience), PE anti-podoplanin (8.1.1 clone; BioLegend), and anti-human/mouse fibronectin (HFN7.1 clone; Novus Biologicals).

Techniques: Control

Journal: Journal of Functional Biomaterials

Article Title: Multi-Composite Bioactive Osteogenic Sponges Featuring Mesenchymal Stem Cells, Platelet-Rich Plasma, Nanoporous Silicon Enclosures, and Peptide Amphiphiles for Rapid Bone Regeneration

doi: 10.3390/jfb2020039

Figure Lengend Snippet: Number of CB MSC attached to bacterial grade (non-adherent) plastic coated with various protein-containing solutions after 1 hour. 10% PRP resulted in the greatest cell retention, while both PRP and PPP were superior to gelatin or fibronectin coatings ( A ). Percentage of cells retained by coated or gelled PCL (gray) and collagen (white) scaffolds after 24 h compared to MSC seeded onto tissue culture plastic. PRP proved to be the most potent coating agent, while PRP and PRP-PA gels retained greater than 90% of loaded cells in PRP-coated collagen scaffolds ( B ). PA gels were significantly greater for cell retention than any of the coatings (δ, p = 3.0 × 10 −7 ). PA gels retained significantly less cells in either scaffold material than the PRP only gel (Ω, p = 0.012) or the PRP + PA gel carrier (Π, p < 0.002). Collagen retained significantly more cells than PCL scaffolds when delivered via PRP only gel (†, p = 0.023) or PRP + PA gel (‡, p = 0.010).

Article Snippet: Recombinant human bone morphogenetic protein-2 (BMP2) and recombinant human fibronectin were purchased from R&D Systems (Minneapolis, MN, U.S.).

Techniques:

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Interleukin-17A induces renal fibrosis through the ERK and Smad signaling pathways.

doi: 10.1016/j.biopha.2019.109741

Figure Lengend Snippet: Fig. 1. Inhibition of fibronectin expression in UUO kidney by blockade of IL-17A. (a ∼d). IL-17A (a & b) and fibronectin expression (c & d) in 7-day post-UUO kidney (b & d) and unobstructed contralateral kidney (control) (a & c) was determined by immunohistochemistry. Faint staining of IL-17A was observed in normal tubular cells of the control kidney, whereas strong staining IL-17A was found in tubular cells of UUO kidney. Fig. 1, Panel a represents relative percentage of IL-17A staining using immunohistochemistry (IHC) of Fig. 1a and b. Fig. 1, Panel b represents relative percentage of fibronectin staining using IHC of Fig. 1c and d. One represented section is demonstrated in figures (a–d). Magnification: a, b, c, d: ×400.

Article Snippet: The catalog number of fibronectin antibody was MAB1918 (R&D Systems) with 1:100 dilutions.

Techniques: Inhibition, Expressing, Control, Immunohistochemistry, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Interleukin-17A induces renal fibrosis through the ERK and Smad signaling pathways.

doi: 10.1016/j.biopha.2019.109741

Figure Lengend Snippet: Fig. 2. Decreased fibronectin expression, ERK1/2 activation, and TGF-β1 expression by blockade of IL-17A function. Mice received UUO for 7 days and extracellular matrix in kidney specimens was stained with Masson's Trichrome (a: the contralateral control kidney; b: the UUO kidney). Mice were injected in- travenously with 100 μg anti-IL-17A receptor antibody through the tail vein at 2 h before UUO and on days 1 and 3 after UUO (c). One represented section is demonstrated in figures. Fig. 2a, Panel a represents relative percentage of fibrosis staining using IHC of Fig. 2a, 2b, and 2c. Kidney parenchymal lysate was subjected to immunoblotting analysis to determine IL-17A and fibronectin expression (d) and p-ERK1/2 and total ERK1/2 levels (e). The density of each protein band was normalized with that of GAPDH, and relative fold changes compared to the control of Fig. 2d are displayed in the Fig. 2d, Panel a (white bars: fibronectin expression; black bars: IL-17A expression) and relative fold changes compared to the control of Fig. 2e are displayed in the Fig. 2e, Panel a. One representative result from three independent experiments is shown (*p < 0.05). TGF-β1 mRNA expression was analyzed by qPCR. TGF-β1 mRNA expression was normalized to GAPDH, and relative fold changes compared to the control are displayed (n = 4 in each group; *p < 0.05) (f). Kidney parenchymal lysate was subjected to immunoblotting analysis to determine TGF-β1 expression (g). Fig. 2g, Panel a represents western blot analysis shown in Fig. 2g. The groupings of gels were cropped from different parts of the same gel. Magnification: a, b, c: ×200.

Article Snippet: The catalog number of fibronectin antibody was MAB1918 (R&D Systems) with 1:100 dilutions.

Techniques: Expressing, Activation Assay, Staining, Control, Injection, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Interleukin-17A induces renal fibrosis through the ERK and Smad signaling pathways.

doi: 10.1016/j.biopha.2019.109741

Figure Lengend Snippet: Fig. 4. IL-17A–induced increase in fibronectin production. (a). HRPTECs were serum-deprived for 24 h and then grown in the presence or absence of different concentrations of IL-17A (5−100 ng/mL) under serum-free conditions for 2 days. Fig. 4a, Panel a represents wester blot analysis shown in Fig. 4a. (b). Cells were treated with 50 ng/mL IL-17A for dif- ferent periods, as indicated. Quantitative analysis of each protein band was displayed in the supplemental Fig. S2. Cell lysates were subjected to immunoblotting analysis for the measurement of fibronectin production. GAPDH expression was used as a loading control. One representative experiment from at least three independent experiments is shown. The groupings of gels were cropped from different parts of the same gel.

Article Snippet: The catalog number of fibronectin antibody was MAB1918 (R&D Systems) with 1:100 dilutions.

Techniques: Western Blot, Expressing, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Interleukin-17A induces renal fibrosis through the ERK and Smad signaling pathways.

doi: 10.1016/j.biopha.2019.109741

Figure Lengend Snippet: Fig. 5. Blockade of the IL-17A–stimulated increase of ERK1/2 activation and fibronectin expression by an ERK1/2 inhibitor. (a). HRPTECs were stimulated with IL-17A (50 ng/mL) under serum-free conditions for the designated periods. (b & c). Cells were pre-treated with different con- centrations of U0126 (2.5−10 μM/mL) in the presence or absence of IL-17A (50 ng/mL) under serum-free conditions for 7.5 min (b) or 48 h (c). ERK1/2 kinase activity was determined by immunoblotting with an anti-p-ERK1/2 antibody, and the same immunoblot membrane was probed again and total ERK1/2 proteins were then determined. GAPDH expression was used as a loading control. Quantitative analysis of these Western blot analyses was displayed in the supplemental Fig. S3∼5. The groupings of gels were cropped from different parts of the different gels of the same specimen.

Article Snippet: The catalog number of fibronectin antibody was MAB1918 (R&D Systems) with 1:100 dilutions.

Techniques: Activation Assay, Expressing, Activity Assay, Western Blot, Membrane, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Interleukin-17A induces renal fibrosis through the ERK and Smad signaling pathways.

doi: 10.1016/j.biopha.2019.109741

Figure Lengend Snippet: Fig. 6. Inhibition of ERK1/2 activation blocks IL- 17A–induced ECM expression in renal fibroblasts. (a). Hs891 T cells were stimulated with different concentrations of IL-17A (5−100 ng/mL) and grown under serum-free condi- tions for 2 days. Cell lysates were subjected to immunoblotting to determine the expression of fibronectin and type I collagen. (b & c). Cells were pre-treated with different concentrations of U0126 (b: 5−10 μM/mL; c: 2.5−10 μM/mL) in the presence or absence of IL-17A (50 ng/mL) under serum-free conditions for 7.5 min (b) or 48 h (c). ERK1/2 kinase activity was de- termined by immunoblotting with an anti-p-ERK1/2 antibody, and the same immunoblot membrane was probed again and total ERK1/2 proteins were then determined. Fibronectin and type I collagen levels in the supernatant were determined by immunoblotting (c). Quantitative analysis of these Western blot analyses was displayed in the supplemental Figs. S6–S8. The groupings of gels were cropped from different parts of the different gels of the same specimen.

Article Snippet: The catalog number of fibronectin antibody was MAB1918 (R&D Systems) with 1:100 dilutions.

Techniques: Inhibition, Activation Assay, Expressing, Western Blot, Activity Assay, Membrane

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Interleukin-17A induces renal fibrosis through the ERK and Smad signaling pathways.

doi: 10.1016/j.biopha.2019.109741

Figure Lengend Snippet: Fig. 7. Inhibition of IL-17A–stimulated fibronectin using a neutralizing anti-TGF-β1 antibody. a. Cells were serum-deprived for 48 h and stimulated with IL-17A (50 ng/mL) for a further 30 min to 4 h (a). TGF-β1 mRNA ex- pression was analyzed by qPCR. TGF-β1 mRNA expression was normalized to GAPDH, and relative fold changes compared to the control are displayed (n = 5 in each group; *p < 0.05). (b). Cells were treated with an anti-TGF-β1 neu- tralizing antibody (100 ng/mL) in the presence or absence of IL-17A (50 ng/ mL) or TGF-β (5 ng/mL, positive control) for 48 h. (c). Cells were pretreated with different concentrations of an anti-TGF-β1 neutralizing antibody (10−100 ng/mL) followed by stimulation with IL-17A (50 ng/mL) for 48 h. Cell lysates were subjected to immunoblotting to determine fibronectin expression. Quantitative analysis of these Western blot analyses was displayed in the sup- plemental Fig. S9∼S10. The groupings of gels were cropped from different parts of the same gel.

Article Snippet: The catalog number of fibronectin antibody was MAB1918 (R&D Systems) with 1:100 dilutions.

Techniques: Inhibition, Expressing, Control, Positive Control, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Interleukin-17A induces renal fibrosis through the ERK and Smad signaling pathways.

doi: 10.1016/j.biopha.2019.109741

Figure Lengend Snippet: Fig. 8. Inhibition of IL-17A–induced fibronectin expression by inhibiting Smad2/3 activation. (a). Cells were stimulated with IL-17A (50 ng/mL) under serum- free conditions for different periods, as indicated. Cell lysates were subjected to immunoblotting to assess p-Smad2 and p-Smad3 levels. (b ∼d). Cells were pretreated with different concentrations of an anti-TGF-β1 neutralizing antibody (10−100 ng/mL) or SB431542 (5 or 10 μM/mL) followed by stimulation with IL-17A (50 ng/ mL) for 24 h. p-Smad2 or fibronectin levels were determined by immunoblotting. (e). Cells were treated with 50 nM/mL p-Smad2 or p-Smad3 siRNA overnight and incubated with 50 ng/mL IL-17A for 48 h. Fig. 8e, Panel a represents western blot analysis shown in Fig. 8e. Cell lysates were subjected to immunoblotting to determine Smad2, Smad3, and fibronectin levels. The presented results are representative of three independent experiments. Quantitative analysis of these Western blot analyses was displayed in the supplemental Fig. S11∼S14. The groupings of gels were cropped from different parts of the different gels of the same specimen.

Article Snippet: The catalog number of fibronectin antibody was MAB1918 (R&D Systems) with 1:100 dilutions.

Techniques: Inhibition, Expressing, Activation Assay, Western Blot, Incubation