c rad peptide  (Biosynth Carbosynth)

Journal: Theranostics

Article Title: Self-assembling peptide hydrogels functionalized with LN- and BDNF- mimicking epitopes synergistically enhance peripheral nerve regeneration

doi: 10.7150/thno.44276

Figure Lengend Snippet: Self-assembling peptides used in this study

Article Snippet: The SAPs RAD (Ac-RADARADARADARADA-NH 2 ), LN-mimicking peptide-modified SAP RAD-IKV (Ac-RADARADARADARADA-GG-IKVAV-NH 2 ), BDNF-mimicking peptide-modified SAP RAD-RGI (Ac-RADARADARADARADA-GG-RGIDKRHWNSQ-NH 2 ) and bifunctional mimicking peptides-modified SAP RAD-IKV-GG-RGI (Ac-RADARADARADARADA-GG-IKVAV-GG-RGIDKRHWNSQ-NH 2 ) (purity > 90%) were custom-synthesized and purified by ChinaPeptides Co., Ltd (Shanghai, China) and Scilight-Peptide Co., Ltd (Beijing, China).

Techniques:

Journal: Theranostics

Article Title: Self-assembling peptide hydrogels functionalized with LN- and BDNF- mimicking epitopes synergistically enhance peripheral nerve regeneration

doi: 10.7150/thno.44276

Figure Lengend Snippet: Self-assembling peptides used in this study

Article Snippet: The SAPs RAD (Ac-RADARADARADARADA-NH 2 ), LN-mimicking peptide-modified SAP RAD-IKV (Ac-RADARADARADARADA-GG-IKVAV-NH 2 ), BDNF-mimicking peptide-modified SAP RAD-RGI (Ac-RADARADARADARADA-GG-RGIDKRHWNSQ-NH 2 ) and bifunctional mimicking peptides-modified SAP RAD-IKV-GG-RGI (Ac-RADARADARADARADA-GG-IKVAV-GG-RGIDKRHWNSQ-NH 2 ) (purity > 90%) were custom-synthesized and purified by ChinaPeptides Co., Ltd (Shanghai, China) and Scilight-Peptide Co., Ltd (Beijing, China).

Techniques:

Journal: Theranostics

Article Title: Self-assembling peptide hydrogels functionalized with LN- and BDNF- mimicking epitopes synergistically enhance peripheral nerve regeneration

doi: 10.7150/thno.44276

Figure Lengend Snippet: Characterization of the SAP solutions and hydrogels. ( A ) Sequences of the functionalized self-assembling peptides. ( B ) Typical CD spectra of RAD, RAD-IKV, RAD-RGI, RAD-IKV-GG-RGI, RAD/IKV, RAD/RGI, RAD/IKV-GG-RGI, and RAD/IKV/RGI solutions. ( C ) AFM images of the peptide solutions and SEM morphologies of the hydrogels. ( D ) Rheological characterization of the peptide hydrogels by strain sweep studies, where the storage modulus (G') and loss storage modulus (G'') were recorded as a function of oscillation strain (%). ( E ) Dynamic frequency sweep test (0.1-10 rad/s at 0.5% strain) of the peptide hydrogels.

Article Snippet: The SAPs RAD (Ac-RADARADARADARADA-NH 2 ), LN-mimicking peptide-modified SAP RAD-IKV (Ac-RADARADARADARADA-GG-IKVAV-NH 2 ), BDNF-mimicking peptide-modified SAP RAD-RGI (Ac-RADARADARADARADA-GG-RGIDKRHWNSQ-NH 2 ) and bifunctional mimicking peptides-modified SAP RAD-IKV-GG-RGI (Ac-RADARADARADARADA-GG-IKVAV-GG-RGIDKRHWNSQ-NH 2 ) (purity > 90%) were custom-synthesized and purified by ChinaPeptides Co., Ltd (Shanghai, China) and Scilight-Peptide Co., Ltd (Beijing, China).

Techniques:

Journal: BMC Cell Biology

Article Title: Actomyosin contractility controls cell surface area of oligodendrocytes

doi: 10.1186/1471-2121-10-71

Figure Lengend Snippet: Integrins in cell surface expansion and endocytosis in Oli-neu cells . (A) Oli-neu cells were treated for 8 h with 100 nM RGD-peptide, 100 nM inactive RAD-peptide, 10 μM Y27632 and 50 μM blebbistatin (blebb) as indicated. Cells were stained with Alexa Fluor 488 conjugated wheat germ agglutinin (green). Scale bar, 10 μm. (B) Changes in relative surface area were quantified by image analysis as described in Material and Methods. (C) Changes in relative surface area shown for cells grown on fibronectin. (D) Quantitative analysis of dextran uptake (30 min) in Oli-neu cells treated with 100 nM RGD-peptide or inactive RAD-peptide for 8 h. Values represent means ± SEM (n > 70 cells, **p < 0,01; ***p < 0,001).

Article Snippet: The effects of RGD-binding integrins were assessed by treating Oli-neu cells for 8 h with 100 nM RGD-peptide (#05231701, Calbiochem) or with the inactive control RAD-peptide (#03340052, Calbiochem).

Techniques: Staining

Journal: BMC Cell Biology

Article Title: Actomyosin contractility controls cell surface area of oligodendrocytes

doi: 10.1186/1471-2121-10-71

Figure Lengend Snippet: Matrix rigidity regulates cell surface area and endocytosis in Oli-neu cells . (A) Quantification of the elastic shear moduli of polyacrylamid gels on glass coverslips with varying amounts of bisacrylamid. Values represent means ± SD (n > 30 measurements). (B) Oli-neu cells were cultured for 1 d on polyacrylamid gels of different rigidities (using bisacrylamid varying from 0,5% to 0,01%) and analyzed for surface area differences by staining with Alexa Flour 488 conjugated wheat germ agglutinin (green). Scale bar, 10 μm. (C) Quantification of the surface area of Oli-neu cells on polyacrylamid gels of different rigidities (mean ± SEM; n > 100 cells, ***p < 0,001). (D+E) Cells were cultured for 1 d on polyacrylamid gels of different rigidities by varying the amount of bisacrylamid. Changes in the amount of dextran uptake ((D) 15 min and (E) 30 min) were quantified. Values represent the mean ± SEM (n > 60 cells from three independent experiments, ***p < 0,001). (F) Quantification of cell surface area of Oli-neu cells cultured on polyacrylamid gels for 1 d and treated for 10 h with 10 μM Y27632 or C3 transferase (mean ± SEM; n > 100 cells, *p < 0,05, **p < 0,01, ***p < 0,001). (G) Quantitative analysis of surface area changes of Oli-neu cells cultured for 1 d on polyacrylamid gels and treated with 50 μM blebbistatin (blebb) for 10 h (mean ± SEM; n > 60 cells, ***p < 0,001). (H) Oli-neu cells were cultured on polyacrylamid gels of different rigidities, treated with RGD-peptide or inactive RAD-peptide and changes in relative surface area were quantified. Values represent the mean ± SEM (n > 80 cells, *p < 0,05, **p < 0,01, ***p < 0,001).

Article Snippet: The effects of RGD-binding integrins were assessed by treating Oli-neu cells for 8 h with 100 nM RGD-peptide (#05231701, Calbiochem) or with the inactive control RAD-peptide (#03340052, Calbiochem).

Techniques: Cell Culture, Staining

Journal: PLoS ONE

Article Title: Integrin – Dependent Mechanotransduction in Mechanically Stimulated Human Annulus Fibrosus Cells: Evidence for an Alternative Mechanotransduction Pathway Operating with Degeneration

doi: 10.1371/journal.pone.0072994

Figure Lengend Snippet: Cells derived from non-degenerate and degenerate IVDs were treated+/− RAD (50 µg/ml) or RGD (50 µg/ml) -peptides 30 minutes prior to mechanical stimulation with CTS at 10% strain, 1.0 Hz, for 20 minutes, then incubated for up to 24 hours prior to analysis. QRT-PCR was used to analyse the gene expression of A) ADAMTS -4 or B) type I collagen, relative to the housekeeping gene GAPDH and normalised to the corresponding unloaded baseline control in non-degenerate (n = 3) and degenerate (n = 3) AF cells, respectively. Black represents AF cells cyclically strained without peptide treatment, while speckles and stripes represent cells cyclically strained after treatment with RAD or RGD –peptides, respectively. Values are mean of 3 donors+/− standard error mean. *denote a significant change ( p ≤0.05) in gene expression between mechanically stimulated and unstimulated baseline control.

Article Snippet: AF cells in serum-free media adhered to Bioflex® culture plates were treated with or without RAD control peptides (50 µg/mL) (Calbiochem, Cat no. 03-34-0052) or the function blocking peptides RGD (50 µg/mL) (Calbiochem, Cat no. 03-34-0035) 30 minutes prior to the application of CTS (peptide concentrations and treatment durations obtained from previous chondrocyte and NP cell studies , ).

Techniques: Derivative Assay, Incubation, Quantitative RT-PCR, Expressing

Journal: PLoS ONE

Article Title: Integrin – Dependent Mechanotransduction in Mechanically Stimulated Human Annulus Fibrosus Cells: Evidence for an Alternative Mechanotransduction Pathway Operating with Degeneration

doi: 10.1371/journal.pone.0072994

Figure Lengend Snippet: AF cells derived from non-degenerate IVDs (n = 4) were treated+/− RGD (50 µg/ml) or RAD (50 µg/ml) peptides, and mechanically stimulated (10% CTS, 1.0 Hz frequency) in serum-free media and total protein extracted at timepoints of 5 and 20 minutes. Mechanically stimulated and unstimulated,+/− RGD or RAD peptides, non-degenerate protein samples (5 µg/well) exposed to A) 5 minutes and B) 20 minutes of CTS, were separated using 10% SDS-PAGE and probed using primary antibodies against phosphorylated FAK. Blots were then stripped using a stripping buffer, re-blocked and probed using an antibody against total FAK protein. C) The density of bands were quantified using a Syngene imaging system and the ratio of phosphorylated: total FAK protein normalised to timepoint controls and plotted as % change. *denotes a significant change ( p ≤0.05) between treatment groups.

Article Snippet: AF cells in serum-free media adhered to Bioflex® culture plates were treated with or without RAD control peptides (50 µg/mL) (Calbiochem, Cat no. 03-34-0052) or the function blocking peptides RGD (50 µg/mL) (Calbiochem, Cat no. 03-34-0035) 30 minutes prior to the application of CTS (peptide concentrations and treatment durations obtained from previous chondrocyte and NP cell studies , ).

Techniques: Derivative Assay, SDS Page, Stripping Membranes, Imaging