Journal: Antioxidants

Article Title: Olive Leaf Extract Added to Losartan Treatment Improved Klotho/Wnt/β-Catenin Signaling in Hypertensive Rats with Focal Segmental Glomerulosclerosis

doi: 10.3390/antiox15010146

Figure Lengend Snippet: Effects of L, L+T and L+O on 6–nitrotryptophan, Klotho, Wnt4, β–catenin, PAI–1 and fibronectin protein expressions in the kidney of SHR with ADR-induced FSGS. The analysis of 6–nitrotryptophan/actin ( A ), Klotho/actin ( B ), Wnt4/GAPDH ( C ), β–catenin/GAPDH ( D ), PAI–1/GAPDH ( E ), fibronectin/GAPDH ( F ) protein expressions and representative Western blots ( G ). Values represent mean ± SEM of three to four independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. SHC; # p < 0.05 and ### p < 0.001 vs. SHADR; $$ p < 0.01 and $$$ p < 0.001 vs. SHADR+L; and &&& p < 0.001 vs. SHADR+L+T. SHC, control group; SHADR, model group; L, losartan; T, tempol; and O, olive leaf extract.

Article Snippet: The membranes were blocked with 5% non-fat dry milk in Tris-buffered saline with 0.1% Tween 20 (TBS-Tween) and then incubated with the following primary antibodies: 6-nitrotryptophan (1:1000, ab243072, Abcam, Cambridge, UK), Klotho (1:1000, NBP1-76511, Novus biologicals, LCC, USA), Wnt-4 (1:2000, NBP2-20909, Novus biologicals, LCC, Centennial, CO, USA), β-catenin (1:300, AF1329, R&D Systems, Inc., Minneapolis, MN, USA), PAI-1 (1:1000, NBP1-19773, Novus biologicals, LCC, Centennial, CO, USA), fibronectin (1:1000, ab2413, Abcam, Cambridge, UK), actin (1:500, A5060, Sigma-Aldrich, MO, USA) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:2500, ab9485, Abcam, Cambridge, UK).

Techniques: Western Blot, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Angiogenic Potential of Tissue Engineered Cartilage From Human Mesenchymal Stem Cells Is Modulated by Indian Hedgehog and Serpin E1

doi: 10.3389/fbioe.2020.00327

Figure Lengend Snippet: Blocking IHH or Serpin E1 increased angiogenic potential of CM of chondrogenically differentiated BMSC in vitro and showed no effect in vivo CAM assay. (A) Quantification of endothelial cell migration assay comparing the impact of pre-incubation of CM with blocking antibodies for factors of interest. Values are normalized to the BMSC-CM condition. UCM depicts the un-conditioned medium. (B) Quantification of endothelial cell proliferation assay comparing the impact of pre-incubation of CM with blocking antibodies for factors of interest. Values are normalized to the BMSC-CM condition. UCM depicts the un-conditioned medium. (C) Images of chick chorioallantoic membrane incubated for 72 h with CM (blocked for 30 min) soaked 5 mm filter disks. Filter circles were placed on a vessel free area on day 7 of the chick development. (D) Boxplot depicting the ranking of the angiogenic potential (lowest = 1 highest = 45: average of 3 independent observers: box = interquartile range, whiskers + 1–99%). N = 5 per condition. A & B were performed in two batches each pooling 3 donors; n = 3. Significance was determined by mixed-linear model with Bonferroni post hoc test.

Article Snippet: Blocking antibodies against VEGF-A (Cat. # AF-293-SP) and Serpin E1 (Cat.# MAB1786-SP, Clone # 242816) were obtained from R&D-Systems (Minneapolis, MN, United States) and utilized at 0.5 μg/mL according to the manufacturers in vitro testing protocol.

Techniques: Blocking Assay, In Vitro, In Vivo, Chick Chorioallantoic Membrane Assay, Cell Migration Assay, Incubation, Proliferation Assay, Membrane