purified recombinant egfp  (Sangon Biotech)

Journal: eLife

Article Title: Axon-specific microtubule regulation drives asymmetric regeneration of sensory neuron axons

doi: 10.7554/eLife.104069

Figure Lengend Snippet:

Article Snippet: To investigate the in vitro formation of the stem axon in DRG neuron development, we transduced these cells with an ultra-purified recombinant AAVPHP.S-CMV-eGFP (#VB010000-9394npt, VectorBuilder) virus at DIV 9 at a concentration of 1.12 × 10 10 cfu/ 7000 cells.

Techniques: Knock-Out, Transfection, Construct, Plasmid Preparation, Transduction, IF-cells, Sequencing, RNAscope, Blocking Assay, Multiplex Assay, Software, Staining, Formulation, Immunofluorescence, Amplification, Live Cell Imaging, Imaging, Western Blot

Journal: Scientific Reports

Article Title: Neurogenic differentiation factor NeuroD confers protection against radiation-induced intestinal injury in mice

doi: 10.1038/srep30180

Figure Lengend Snippet: ( A ) Purified NeuroD-EGFP fusion proteins were analyzed by 12% SDS-PAGE and stained with Coomassie Blue. Lanes are as follows: lane 1, supernatant after sonication; lane 2, effluent after loading; lane 3, 100 mM imidazole elution buffer; lane 4, 200 mM imidazole elution buffer; lane 5, 300 mM imidazole elution buffer; lane M, protein marker. (B) Purified NeuroD-EGFP fusion proteins were analyzed by Western blot. (C) Transduction of NeuroD-EGFP into IEC-6 rat jejunal crypt cell line. Nuclear DNA (blue) was stained with DAPI. Scale bar: 20 μm. (D) Transduction of NeuroD-EGFP into mouse small intestine (400x magnification).

Article Snippet: After a 24-h incubation, the medium was replaced by fresh medium containing 1 μM of purified recombinant EGFP (Sangon Biotech, Shanghai, China) or NeuroD-EGFP protein.

Techniques: Purification, SDS Page, Staining, Sonication, Marker, Western Blot, Transduction

Journal: Scientific Reports

Article Title: Neurogenic differentiation factor NeuroD confers protection against radiation-induced intestinal injury in mice

doi: 10.1038/srep30180

Figure Lengend Snippet: ( A ) Kaplan-Meier survival analyses of C57BL/6J mice after 8 Gy TBI with or without NeuroD-EGFP treatment. (B) Mean survival time of dead animals in each group. * P < 0.05; *** P < 0.001. (C) Body weight change of C57BL/6J mice after 8 Gy TBI with or without NeuroD-EGFP treatment.

Article Snippet: After a 24-h incubation, the medium was replaced by fresh medium containing 1 μM of purified recombinant EGFP (Sangon Biotech, Shanghai, China) or NeuroD-EGFP protein.

Techniques:

Journal: Scientific Reports

Article Title: Neurogenic differentiation factor NeuroD confers protection against radiation-induced intestinal injury in mice

doi: 10.1038/srep30180

Figure Lengend Snippet: (A) Representative H&E staining of intestinal sections from PBS, EGFP or NeuroD-EGFP treated mice at 3.5 days after 9 Gy TBI (100x or 200x magnification). Bar graph of villus height (B) , crypt depth (C) , and crypts per circumference (D) determined by measuring vertically well-oriented crypt villus units from H&E-stained sections of mice after 9 Gy TBI. Normal, sham-irradiated control. For villus height or crypt depth measurement, at least 30 villi or crypts per mouse were measured. Crypts per circumference were counted in 3 separate tubular intestinal slices for each mouse. * P < 0.05; ** P < 0.01; *** P < 0.001; n = 3.

Article Snippet: After a 24-h incubation, the medium was replaced by fresh medium containing 1 μM of purified recombinant EGFP (Sangon Biotech, Shanghai, China) or NeuroD-EGFP protein.

Techniques: Staining, Irradiation, Control

Journal: Scientific Reports

Article Title: Neurogenic differentiation factor NeuroD confers protection against radiation-induced intestinal injury in mice

doi: 10.1038/srep30180

Figure Lengend Snippet: (A) Representative Ki67-immunostained intestinal sections from PBS, EGFP or NeuroD-EGFP treated mice at 3.5 days after 9 Gy TBI (200x or 400x magnification). (B) Quantification of Ki67-positive cells per crypt determined from panel (A). At least 30 well-oriented crypts per mouse were counted. *** P < 0.001; n = 3. (C) Representative TUNEL-staining of intestinal sections from PBS, EGFP or NeuroD-EGFP treated mice at 6 hours after 9 Gy TBI. TUNEL-stained epithelium shows green fluorescence. Nuclear DNA (blue) was stained with DAPI. Scale bar: 25 μm. (D) Quantification of TUNEL-positive cells determined from panel (C). The average number of positive cells in ten fields (400x magnification) per treated mouse was determined. * P < 0.05; n = 3.

Article Snippet: After a 24-h incubation, the medium was replaced by fresh medium containing 1 μM of purified recombinant EGFP (Sangon Biotech, Shanghai, China) or NeuroD-EGFP protein.

Techniques: TUNEL Assay, Staining, Fluorescence

Journal: Scientific Reports

Article Title: Neurogenic differentiation factor NeuroD confers protection against radiation-induced intestinal injury in mice

doi: 10.1038/srep30180

Figure Lengend Snippet: (A) Map of gene clusters for those differentially expressed between EGFP and NeuroD-EGFP treated mice at 12 hours post 9 Gy TBI. (B) Volcano plot comparing EGFP treatment versus NeuroD-EGFP treatment. Genes with fold change ≥2 and P value ≤ 0.05 are marked with red dots, while those with fold change ≤−2 and P value ≤ 0.05 are marked with green dots. (C) Significant pathways altered in mice 12 hours post 9 Gy TBI (EGFP treated mice vs . NeuroD-EGFP treated mice). (D) Quantitative real-time PCR validation for differentially expressed genes between EGFP and NeuroD-EGFP treated mice. * P < 0.05; ** P < 0.01; n = 3. ( E ) NeuroD-EGFP induced TIMP-1 expression. The expression of TIMP-1 in the mice small intestine was determined by immunohistochemical staining with a TIMP-1 antibody (200x or 400x magnification).

Article Snippet: After a 24-h incubation, the medium was replaced by fresh medium containing 1 μM of purified recombinant EGFP (Sangon Biotech, Shanghai, China) or NeuroD-EGFP protein.

Techniques: Real-time Polymerase Chain Reaction, Biomarker Discovery, Expressing, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: Neurogenic differentiation factor NeuroD confers protection against radiation-induced intestinal injury in mice

doi: 10.1038/srep30180

Figure Lengend Snippet: Top 10 genes dysregulated within NeuroD-EGFP- and EGFP-treated mice from a 45,200-transcript microarray of small intestinal tissues (NeuroD-EGFP-treated vs. EGFP-treated mice).

Article Snippet: After a 24-h incubation, the medium was replaced by fresh medium containing 1 μM of purified recombinant EGFP (Sangon Biotech, Shanghai, China) or NeuroD-EGFP protein.

Techniques: Microarray, Sequencing