Journal: bioRxiv

Article Title: Physiological perfusion of human vasculature reveals a YAP/TAZ-Apelin switch linking intraluminal flow to endothelial state transitions and vessel remodeling

doi: 10.64898/2026.03.21.713033

Figure Lengend Snippet: (A) T1 endothelial cells on Day 4 plotted for APLN and APLNR expressions. (B) Rank plot of all analyzed genes and their correlation coefficient with APLN (left). Correlation of APLN expression with gene set scoring for YAP/TAZ targets (center). Overlay of scores onto the scatter plot of APLN and APLNR expressions (right). Expression values are from T1 endothelial cells on Day 4. (C) qPCR expression data of APLN and APLNR from monolayer endothelial cells treated with IAG933 (2 µM; 24 hrs; 3 independent experiments; 2-tailed unpaired student’s t-test). (D, E) Mapping tip cell and stalk-like clusters from the human tumor angiogenesis atlas of Goveia et al . onto T1 endothelial cells on Day 4, with (E) additionally showing RNA velocity streamlines. (F) Gene set scoring of T1 endothelial cells on Day 4 for Apelin up-regulated genes (Park et al ., ). (G) Negative correlation between gene set scores for YAP/TAZ targets and Apelin up-regulated genes. (H) Box plot summaries of (G). (A, B, G) Spearman’s correlation coefficient. (A, B, D-F) Axes show log-normalized expression values of APLN and APLNR denoised using Markov Affinity-based Graph Imputation of Cells. (I) Skeleton visualizations of vascular beds grown using low or high flow on Day 4. (J) Quantification of (I) (11 devices per condition across 6 independent experiments). Data is a subset of (L) and (N). (K) Skeleton visualizations of low/high flow vascular beds on Day 4 treated with IAG933 (2 µM). (L) Quantification of (K) (6 devices per condition across 3 independent experiments). (M) Skeleton visualizations of low/high flow vascular beds on Day 4 treated with ML221 (10 µM). (N) Quantification of (M) (5-6 devices per condition across 3 independent experiments). (J, L, N) 2-tailed unpaired Welch’s t-test. (C, J, L, N) * p < 0.05; ** p < 0.01; *** p < 0.001. Error bars show SEM.

Article Snippet: IAG933 (Piramal Pharma Solutions, kindly provided by Methvin Isaac, Ontario Institute for Cancer Research) was used at 2 μM, ML221 (MedChemExpress) was used at 10 μM, human recombinant BMP (R&D Systems) was used at 10 ng/mL, and Axitinib (MedChemExpress) was used at 100 nM.

Techniques: Expressing

Journal: Nature Communications

Article Title: A temperature-regulated circuit for feeding behavior

doi: 10.1038/s41467-022-31917-w

Figure Lengend Snippet: a Schematic image of injecting AAVretro-GFP into PVH or ARC, and GFP-labeled cell body sucked by a micropipette. Scale bar, 20 µm. b Volcano plot and bar graph depicting the transcript abundance between PVH-projecting (red) and ARC-projecting (green) apMPOA neurons. c Venn diagram depicting the shared and unique gene expressions of the two neuronal populations. The expression (FPKM) of top ten DEGs in the KEGG pathway categories: metabolism ( d ), environmental information processing ( e ), organismal systems ( f ), cellular processes ( g ). The candidate marker genes are indicated in red #. n = 5 animals for each group. h The genes related to feeding peptides and receptors. The candidate marker genes are indicated in red #. n = 5 animals for each group. i Schematic and representative images of cannula injection of antagonists or agonists of the candidate genes. Scale bar, 100 µm. j Injection of antagonists or agonists of the selected candidate genes during normothermia shows that apelin, galanin, and NPY injections cause food intake changes. One-way repeated-measure ANOVA and post hoc LSD multiple comparison, *** p < 0.001, ** p = 0.01. n = 14 animals. k Feeding test after apelin, galanin and NPY injections. Compared with PBS, apelin-13 robustly suppressed food intake, and the galanin robustly promoted food intake at ambient temperatures. NPY only promoted food intake during normothermia, but not during high or low temperature. Two-way repeated-measure ANOVA and post hoc LSD multiple comparison. Interaction effect: F 6,96 (temperature × drug) = 6.03, *** p < 0.001; main effect: F 2,47 (temperature) = 29.81, *** p < 0.001; F 3,48 (drug) = 28.62, *** p < 0.001. n = 14 animals. l Feeding test after apelin, galanin, and NPY receptors antagonists injections. Compared with PBS, ML221 (apelin receptor antagonist) and M40 (GalR1 antagonist) reversed the food intake by the agonists. Blocking the NPY receptor (BIBO 3304) did not induce food intake changes. Two-way repeated-measure ANOVA and post hoc LSD multiple comparison. Interaction effect: F 6,100 (temperature × drug) = 2.4, * p = 0.033; main effect: F 2,49 (temperature) = 29.81, *** p < 0.001; F 3,50 (drug) = 13.43, *** p < 0.001. n = 14 animals. m Colocalizations of GalR1 (green), ARC-projecting apMPOA neurons (red), and fasting-induced C-Fos (purple). Image representative of n = 5 mice. Scale bar, 100 µm. Arrowhead denotes GalR1 +, ARC-projecting+, and C-Fos+ neurons, asterisk denotes GalR1 + and ARC-projecting+ neurons. n ARC-projecting neurons presented colocalization with fasting-induced C-Fos, and galanin receptor 1. o Colocalizations of Aplnr (green), PVH-projecting apMPOA neurons (red), and fasting-induced C-Fos (purple). Image representative of n = 5 mice. Scale bar, 100 µm. Arrowhead denotes Aplnr +, PVH-projecting+, and C-Fos+ neurons, asterisk denotes Aplnr + and PVH-projecting+ neurons. p PVH-projecting neurons presented colocalizations with apelin receptor, but rarely with fasting-induced C-Fos. All error bars show SEM.

Article Snippet: Fifteen min after bilateral injections, the mice were returned to their home cages and feeding behavior tests were performed for 2 h. The drugs were prepared fresh on the day of the experiment by dilution in vehicle solution (PBS supplemented with 0.1% DMSO, 5% PEG400, and 5% Tween-80), including the integrin inhibitor SB273005 (Selleck, 150 nL/unilateral, 1 μM), the enolase inhibitor AP-III-a4 (ENOblock, Selleck, 150 nL/unilateral, 1 μM), the IRAK4 inhibitor PF-06650833 (Selleck, 150 nL/unilateral, 1 μM), the antagonist of apelin receptor ML221 (Selleck, 150 nL/unilateral, 1 μM), the serotonin reuptake inhibitor Fluvoxamine maleate (MK-264, Selleck, 150 nL/unilateral, 1 μM), the antagonist of dopamine D3 receptor BP897 (Adooq Bioscience, 150 nL/unilateral, 1 μM), the antagonist of NPY Y1 receptor BIBO 3304 trifluoroacetate (Tocris, 150 nL/unilateral, 1 μM) and antagonist of galanin receptor M40 (Tocris, 150nL/unilateral, 1 μM).

Techniques: Labeling, Expressing, Marker, Injection, Comparison, Blocking Assay

Journal: Development (Cambridge, England)

Article Title: Thymic epithelial organoids mediate T-cell development.

doi: 10.1242/dev.202853

Figure Lengend Snippet: Fig. 2. TECs cultured as organoids show in vitro functionality when reaggregated with T-cell progenitors. (A) Workflow to generate organoid reaggregate fetal thymic organ cultures (ORFTOCs). (B) Flow cytometry plot showing T-cell development in D6 ORFTOCs. (C) Proportion of DP thymocytes at D6 in ORFTOCs and controls [fetal thymic organ cultures (FTOCs) and reaggregates containing only MEFs and the EpCAM-depleted cells from thymic lobes]. **P=0.0047; ns, not significant (P>0.05) (Kruskal–Wallis test with Dunn’s multiple comparison test; n=9, 16 and 9 for ORFTOCs, FTOCs and reaggregates from MEFs+EpCAM-depleted cells, respectively, from 9 independent experiments). (D,E) Flow cytometry plots. (D) T-cell development at D6 in controls (left: FTOCs; right: reaggregates from MEFs+EpCAM-depleted cells). (E) Absence of a CD45– Lineage−(Lin−) population in control reaggregates containing only TECs cultured as organoids and MEFs. Gating strategy is indicated on the left for all flow cytometry plots. (F) Proportion of CD3ɛ-positive, TCRβ-positive cells in ORFTOCs and FTOCs at D6, D13 and D20. **P=0.0021, ***P=0.0003, ****P<0.0001; ns, not significant (P>0.05) (two-way ANOVA with Tukey’s multiple comparison test; n=9, 6, 3, 16, 8 and 5 for D6 ORFTOCs, D13 ORFTOCs, D20 ORFTOCs, D6 FTOCs, D13 FTOCs and D20 FTOCs, respectively, from at least 3 independent experiments). All graphs represent individual datapoints with mean±s.d. (G) RNAscope for Foxn1 in a D13 ORFTOC. Nuclei are counterstained using Spectral DAPI (grey). (H-J) Immunofluorescence images. (H) KRT8 (blue) and MHCII (green) staining in a D13 ORFTOC (left) and a D13 FTOC (right). (I) Medullary cells (UEA1 reactivity; magenta) in a D13 ORFTOC. (J) Epithelial cells (EpCAM; blue), medullary cells (UEA1 reactivity; bright pink) and T cells (CD3ɛ; amber) in a D13 ORFTOC. DAPI stains nuclei (grey). ORFTOCs were formed with TECs cultured as organoids originating from E16.5 thymi, with the EpCAM-depleted fraction of cells from E13.5 thymi and with MEFs. FTOCs were from E13.5 thymi. Scale bars: 100 µm (G-I); 10 μm (J).

Article Snippet: The cells were incubated for 20 min with the following antibodies: Ter119-FITC (BioLegend, 116205; 1/800), Cd45R-FITC (BioLegend, 103205; 1/800), CD11b-FITC (Thermo Fisher Scientific, 11-0112-82; 1/800), Ly-6G-FITC (BioLegend, 108405; 1/800), Cd11C-FITC (BioLegend, 117306; 1/800), NK-1.1-FITC (BioLegend, 108705; 1/800) (together referred to as Lineage), CD44-PE (BioLegend, 103008; 1/160), CD69-APC (BioLegend, 104513; 1/160), CD4-BV605 (BioLegend, 100548; 1/40), CD3-PerCP/Cy5.5 (BioLegend, 100327; 1/160), CD8-PE/Cy7 (BioLegend, 100722; 1/160), CD25-BV711 (BioLegend, 102049; 1/160), CD45-AF700 (BioLegend, 103127; 1/160), TCRβ-BV421 (BioLegend, 109230; 1/80) and DAPI (Tocris, 4748, 0.5 μg/ml).

Techniques: Cell Culture, In Vitro, Flow Cytometry, Comparison, Control, RNAscope, Immunofluorescence, Staining

Journal: BMC Ophthalmology

Article Title: Protective effect of apelin-13 in lens epithelial cells via inhibiting oxidative stress-induced apoptosis

doi: 10.1186/s12886-024-03746-6

Figure Lengend Snippet: Apelin-13 prevented H 2 O 2 -induced cellular viability in HLE-B3 cells. Viability of HLE-B3 cells as assessed using CCK8 assay. HLE-B3 cells were treated with ML221 (10 µmol) for 1 h, preincubated with apelin-13 (0.1 µM) for 24 h, and subsequently treated with H 2 O 2 (200 µM) for 24 h in this experiment. Representative images of three replicated experiments were presented. The significance marked at the top of the columns refers to comparison with the control group. Data are shown as mean ± SD ( n = 3)

Article Snippet: HLEC-B3 cells were preincubated with different concentrations of apelin-13 (MCE, New Jersey, USA) and/or ML221 (Anaspec Inc, CA, USA) for 24 h, then treated with H 2 O 2 (KEHBIO, China).

Techniques: CCK-8 Assay, Comparison, Control