vegfa detection  (ProSci Incorporated)

Journal: Development (Cambridge, England)

Article Title: A null mutation of Hhex results in abnormal cardiac development, defective vasculogenesis and elevated Vegfa levels.

doi: 10.1242/dev.01393

Figure Lengend Snippet: Fig. 8. The spatial expression pattern of Vegfa expression is not altered in Hhex–/– embryos. (A) and (B) Low-power views of sagittal sections of E9.5 embryos stained with Hoecsht (blue staining of cell nuclei). Higher-power views of the areas boxed in (A) and (B) are shown as indicated. Green staining represents presence of Vegfa. In the heart of both Hhex+/+ (C) and Hhex–/– (D) embryos, Vegfa is expressed in both the myocardium and endocardium. Vegfa is also highly expressed in the developing gut endoderm of both Hhex+/+ (E) and Hhex–/– (F) embryos.

Article Snippet: Whole hearts were isolated from E9.5-E13.5 Hhex+/+ and Hhex–/– embryos, sonicated in PBS and Vegfa levels were assayed using the mouse Vegfa ELISA detection kit QuantikineM (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Staining

Journal: Poultry Science

Article Title: Ligusticum chuanxiong promotes the angiogenesis of preovulatory follicles (F1–F3) in late-phase laying hens

doi: 10.1016/j.psj.2022.102430

Figure Lengend Snippet: Compounds characterization of CX extract and dose-dependent effects of CX extract on angiogenesis of preovulatory follicles. Based peak intensity chromatograms of CX extract acquired by LC-MS, (A) positive mode and negative (B). (C) Blood vessel was observed on surface of preovulatory follicles (F1–F3) after CX extract treated. (D) Number of follicles in each group after treatment with CX extract. (E) HE staining to detect the granulosa layers thickness after CX extract treated. Scale bar = 100μm. (F) Histogram of granulosa layer thickness ratio. (G) Elisa analysis of VEGFA concentration in granulosa layer (F1–F3) after treatment by CX extract. Immunofluorescence staining of CD31 in CX extract-treated preovulatory follicles, (H) F1, (I) F2, and (J) F3. Scale bar = 100μm. All experiments were performed in triplicate, and the data are the mean ± S.E.M (* P < 0.05, ⁎⁎ P < 0.01, ⁎⁎⁎ P < 0.001).

Article Snippet: After blocking, membranes were incubated with primary antibodies for the detection of CD31 (1:1000), VEGFA (1:500), VEGFR2 (1:1000), P-VEGFR2 (1:500), HIF1-α (1:1000), p-PI3K (1:500), PI3K (1:1000), p-AKT (1:500), AKT (1:1000), p-MTOR (1:500), MTOR (1:500), p-P70S6K (1:500), P70S6K (1:500), RAS (1:500), RAF (1:500), p-MEK (1:500), MEK (1:500), p-ERK (1:500), ERK (1:500), MMP2 (1:500), MMP9 (5:100), and β-actin (1:1000) and secondary antibodies (ABclonal, Wuhan, China).

Techniques: Liquid Chromatography with Mass Spectroscopy, Staining, Enzyme-linked Immunosorbent Assay, Concentration Assay, Immunofluorescence

Journal: Poultry Science

Article Title: Ligusticum chuanxiong promotes the angiogenesis of preovulatory follicles (F1–F3) in late-phase laying hens

doi: 10.1016/j.psj.2022.102430

Figure Lengend Snippet: Dose-dependent effects of CX extract on theca layers in preovulatory follicles of late-phase laying hens. (A), (B), and (C) Western blot to detect the expression of P-VEGFR2, VEGFR2 and CD31 in F1 (A), F2 (B) and F3 (C) theca layers after CX extract treated, quantified according to the western blot. β-actin was included as a loading control. (D), (E), and (F) Immunofluorescence staining of P-VEGFR2 in CX extract-treated F1 (D), F2 (E), and F3 (F). Scale bar = 100 μm. (G), (H), and (I) Immunofluorescence staining of VEGFR2 in CX extract-treated F1 (G), F2 (H), and F3 (I). Scale bar = 100 μm. All experiments were performed in triplicate, and the data are the mean ± S.E.M (* P < 0.05, ⁎⁎ P < 0.01, ⁎⁎⁎ P < 0.001).

Article Snippet: After blocking, membranes were incubated with primary antibodies for the detection of CD31 (1:1000), VEGFA (1:500), VEGFR2 (1:1000), P-VEGFR2 (1:500), HIF1-α (1:1000), p-PI3K (1:500), PI3K (1:1000), p-AKT (1:500), AKT (1:1000), p-MTOR (1:500), MTOR (1:500), p-P70S6K (1:500), P70S6K (1:500), RAS (1:500), RAF (1:500), p-MEK (1:500), MEK (1:500), p-ERK (1:500), ERK (1:500), MMP2 (1:500), MMP9 (5:100), and β-actin (1:1000) and secondary antibodies (ABclonal, Wuhan, China).

Techniques: Western Blot, Expressing, Control, Immunofluorescence, Staining