Journal: Journal of nanobiotechnology

Article Title: ALPL regulates pro-angiogenic capacity of mesenchymal stem cells through ATP-P2X7 axis controlled exosomes secretion.

doi: 10.1186/s12951-024-02396-6

Figure Lengend Snippet: Fig. 5 ALPL mutation increases hBMMSCs exosomes secretion through ATP axis. (A) HPP hBMMSCs was treated with 2 U/mL ATP-apyrase (HPP + Apy) and the exosomes markers were analyzed. Exosomes proteins from equal volumes of culture supernatant of Nor, HPP and HPP + Apy hBMMSCs were loaded for Western Blot. (B) Exosomes volumes derived from each groups were detected by exosomes ELISA complete kit. (C-D) The intracellular expression of CD9 and CD81 were analyzed by Western Blot. (E) Immunofluorescent staining showed CD9 and CD81-positive labeled exosomal proteins localized in Nor, HPP and HPP + Apy hBMMSCs. Scale bar, 10 μm. (F) Nor hBMMSCs was treated with 10µmol/L ATP and the exosomes markers were analyzed. Exo somal proteins from equal volumes of culture supernatant of Nor and Nor + ATP hBMMSCs were loaded for Western Blot. (G) Exosomes volumes derived from Nor and Nor + ATP groups were detected by exosomes ELSA complete kit. (H-I) The intracellular expression of CD9 and CD81 were analyzed by Western Blot. (J) CD9 and CD81-positive labeled exosomal proteins localized in Nor and Nor + ATP hBMMSCs, as assessed by immunofluorescent staining. Scale bar, 10 μm. All results were generated in three independent experiments. Data were shown as mean ± standard deviation (SD); *P < 0.05; **P < 0.01; ***P < 0.001

Article Snippet: The secreted VEGF, PDGFBB, Angiostatin and Endostatin in the samples were quantified using human VEGF ELISA Kit (Proteintech, KE00216, China), human PDGFBB ELISA Kit (Proteintech, KE00161, China), human Angiostatin ELISA kit (Neobioscience Technology, E-EL-H6165, China) and human Endostatin ELISA Kit (Proteintech, KE00259, China) according to manufacture’s instruction.

Techniques: Mutagenesis, Western Blot, Derivative Assay, Enzyme-linked Immunosorbent Assay, Expressing, Staining, Labeling, Generated, Standard Deviation

Journal: Acta physiologica (Oxford, England)

Article Title: Early growth response 1 exacerbates thoracic aortic aneurysm and dissection of mice by inducing the phenotypic switching of vascular smooth muscle cell through the activation of Krüppel-like factor 5.

doi: 10.1111/apha.14237

Figure Lengend Snippet: FIGURE 4 Egr1 knockdown inhibited the PDGF-BB-induced phenotypic switching and migration of VSMCs. (A) The relative mRNA expression level of Egr1 normalized to β-actin in A7R5 cells transfected with pLenti-shEgr1 or pLenti-shRNA (n = 3). (B) Western blot images for protein expression of Egr1, α-SMA, SM22α, OPN, MMP2, and MMP9 in per cell group. (C) Protein expression of Egr1, α-SMA, SM22α, OPN, MMP2, and MMP9 relative to that of β-actin (n = 3). (D) Wound-healing assays were performed to assess the migratory capacity of A7R5s in each group (n = 3) (scale bar = 200 μm). (E) Quantification of the rate of migration in each group via wound-healing assay (n = 3). All data are presented as the mean ± SEM.

Article Snippet: Cells in the PDGF- BB group were cultured with 20 ng/mL PDGF- BB (HY- P7278, MedChemExpress, New Jersey, USA) for 24 h,39,40 whereas those in the control group were treated with PBS.

Techniques: Knockdown, Migration, Expressing, Transfection, shRNA, Western Blot, Wound Healing Assay

Journal: Acta physiologica (Oxford, England)

Article Title: Early growth response 1 exacerbates thoracic aortic aneurysm and dissection of mice by inducing the phenotypic switching of vascular smooth muscle cell through the activation of Krüppel-like factor 5.

doi: 10.1111/apha.14237

Figure Lengend Snippet: FIGURE 5 Egr1 knockdown inhibited the PDGF-BB-induced migration and proliferation of VSMCs. (A) Transwell assays were performed to assess the migratory capacity of A7R5s in each group (n = 3) (scale bar = 100 μm). (B) Quantification of the number of migrated cells in each group via transwell assay (n = 3). (C) Representative images of phalloidin staining for labeling F-actin in cells from each group (scale bar = 50 μm). (D) Analysis of the F-actin area and cell surface area in each group (n = 3). (E) Representative images of ki-67 (red) immunofluorescence staining. Nuclei were stained with DAPI (blue) (scale bar = 100 μm). (F) Quantification of ki-67-positive cells (n = 3). (G) Cell proliferation was detected via cell counting kit-8 (CCK-8) assay at 24, 48, and 72 h after treatment (n = 3). All data are presented as the mean ± SEM.

Article Snippet: Cells in the PDGF- BB group were cultured with 20 ng/mL PDGF- BB (HY- P7278, MedChemExpress, New Jersey, USA) for 24 h,39,40 whereas those in the control group were treated with PBS.

Techniques: Knockdown, Migration, Transwell Assay, Staining, Labeling, Immunofluorescence, Cell Counting, CCK-8 Assay