Journal: The ocular surface

Article Title: Melanocytes as emerging key players in niche regulation of limbal epithelial stem cells.

doi: 10.1016/j.jtos.2021.08.006

Figure Lengend Snippet: Fig. 8. Limbal melanocytes in angiogenesis. A. Immunofluorescence staining of cor neoscleral tissue sections showing Melan-A+ melanocytes and blood vessels in the limbal stroma stained with antibodies against CD31. B. Human vascular endothelial cells (HUVEC) were either untreated (control, low) or treated with angiogenic growth fac tors (AGF, positive control), AGF and IFN-γ (100 U/ml, negative control), or AGF with conditioned media (CM) derived from limbal epithelial stem/progenitor cells (LEPC), limbal mesenchymal stromal cells (LMSC), and limbal melanocytes (LMel). Cell numbers were counted after 6 days. Experi ments were performed in triplicates, and data are expressed as means ± SD (n = 5). (***p = 0.0003; ****p < 0.0001; Mann- Whitney U test). C. HUVEC were either un treated (control, low) or treated with AGF (positive control), AGF and IFN-γ (negative control) (left), or AGF with CM derived from LEPC, LMSC and LMel (right). Cell migration was analyzed by measuring AGF-induced closure of a scratch wound. Experiments were performed in triplicates and data are expressed as means ± SD (n = 3). (*p < 0.05; unpaired two-tailed Student’s t-test). D. Phase contrast images of HUVECs cultured for 24 h on Matrigel in the absence (control, low) or presence of AGF (control) or AGF with CM derived from LEPC, LMSC and LMel (upper panel). The number of nodes (branch sites) (left) and meshes (loops) (right) was quantified as measure of in vitro angiogenesis. Experiments were per formed in triplicate, and data are expressed as means ± SD (n = 10). (*p < 0.05; Mann- Whitney U test).

Article Snippet: On the next day (day 1) and on day 3, the cells were stimulated with CM diluted 1:2 in low ECGM in combination with angiogenic growth factors (AGF), which combined 10 ng/ml vascular endothelial growth factor (VEGF; R&D Systems, Minneapolis, MN) and 10 ng/ml basic fibroblast growth factor (bFGF; Roche Diagnostics).

Techniques: Immunofluorescence, Staining, Control, Positive Control, Negative Control, Derivative Assay, MANN-WHITNEY, Migration, Two Tailed Test, Cell Culture, In Vitro

Journal: Non-Coding RNA

Article Title: Extracellular Small RNAs in Human Milk: Molecular Profiles, Stability and Fragment-Specific Responses in Cell-Based Assays

doi: 10.3390/ncrna12010005

Figure Lengend Snippet: Functional effects of ex-sRNA candidates on HUVEC survival under stress conditions. ( A ) Effect of starvation and exRNA candidates on HUVEC cell survival. The tested exRNA candidates include 5′ tDR-G, 5′ tDR-E, 5′ tDR-V, 5′ tDR-K, 5′ yDR-4, and 5′ rDR-28S. Statistical significance was determined using a one-tailed t -test (Starvation vs. Treated), with significance levels indicated as p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****). ( B ) Effect of VEGF deprivation and exRNA candidates on 0.1% FBS cultured-HUVEC cell survival. tDD accounts to DNA oligonucleotides containing predominantly identical sequence motifs to the exRNA candidate tested within each graph. Statistical significance was determined using ANOVA, with significance levels indicated as p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: HUVEC cells were cultured in RPMI 1640 medium supplemented with 15% FBS, 10 ng/mL VEGF (293-VE-010, R&D system) and 100 units/mL penicillin and streptomycin.

Techniques: Functional Assay, One-tailed Test, Cell Culture, Sequencing

Journal: Endocrinology

Article Title: Androgens Stimulate EPC-Mediated Neovascularization and Are Associated with Increased Coronary Collateralization.

doi: 10.1210/endocr/bqaa043

Figure Lengend Snippet: Figure 3. Androgen treatments increase surface KDR expression and VEGF-induced Akt activation. DHT-treated EEPCs (A–C) and OECs (D–F) were analyzed for cell surface KDR by flowcytometry (A and D); KDR phosphorylation by Western blot (B and E). Akt phosphorylation was assessed in DHT treated EEPCs (C) and OECs (F) following 3 minutes acute stimulation of 10 ng/mL VEGF165. Each experiment was performed in triplicate with EPCs from ≥3 individual donors. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. vehicle control. One-way ANOVA.

Article Snippet: Recombinant VEGF165 (10 ng/mL; R&D Systems) (27) was used to stimulate cell migration.

Techniques: Expressing, Activation Assay, Phospho-proteomics, Western Blot, Control