Fig 1: IGF-1 and RELN as potential mediators in the crosstalk between LECs and cardiomyocytes. (A) Western blot for IGF-1 in heart tissues from control and swim mice (n = 6); (B) RT-qPCR for IGF-1 and RELN in heart tissues from control and swim mice (n = 6); (C) Western blot for IGF-1 in heart tissues from control and swim mice treated with SAR131675 (100 mg/kg/d) or vehicle controls (n = 6); (D) RT-qPCR for IGF-1 and RELN in heart tissues from control and swim mice treated with SAR131675 (100 mg/kg/d) or vehicle controls (n = 6); and ELISA for (E) IGF-1 and (F) RELN levels in LEC-conditioned medium pre-treated with VEGFR3 inhibitor SAR131675 (100 nM) or not (n = 4). Independent-sample t test (two-tailed) was used for statistical comparisons between 2 groups. One-way ANOVA test was used for statistical comparisons among 3 groups. Two-way ANOVA followed by Tukey post hoc test was used for statistical comparisons among 4 groups. * p < 0.05; ** p < 0.01; *** p < 0.001. ANOVA = analysis of variance; GAPDH = glyceraldehyde-3-phosphate dehydrogenase; IGF-1 = insulin-like growth factor-1; LEC = lympathic endothelial cell; OD = optical density; RELN = the extracellular protein Reelin; RT-qPCR = reverse transcription quantitative polymerase chain reaction.
Fig 2: Cardiac lymphangiogenesis is required for exercise-induced physiological cardiac growth by VEGFR3 activation. LEC-conditioned medium promotes both physiological hypertrophy and proliferation of cardiomyocytes through AKT activation and the C/EBPß–CITED4 axis. AKT = protein kinase B; C/EBPß = CCAAT enhancer-binding protein beta; CITED4 = CBP/p300-interacting transactivators with E (glutamic acid)/D (aspartic acid)-rich-carboxylterminal domain4; IGF-1 = insulin-like growth factor-1; LEC = lymphatic endothelial cell; LYVE-1 = lymphatic vessel endothelial hyaluronic acid receptor 1; RELN = the extracellular protein Reelin; VEGFR3 = vascular endothelial growth factor receptor 3.
Fig 3: Harnessing the AIMS-enhanced MSC secretome for immunomodulation applications.a Heatmap measuring the inflammatory factors secreted from IFN-γ primed MSCs from different groups. b–i ELISA assay measuring the secretion of FGF-2, IGF-1, VEGF, HGF-1, IL-6, IL-10, IFN-γ, and TNF- α from different groups after incubating for 4 days. j Flow cytometry analysis measuring the percentage of CD86 and CD163 positive human THP-1 cells after incubating with the conditional medium from MSCs for 2 days. k Flow cytometry analysis measuring the percentage of CD4 positive T cells after incubating with the conditional medium from MSCs for 3 days. l Flow cytometry analysis measuring the B cell proliferation after incubating with the conditional medium from MSCs for 7 days. m, n ELISA assay measuring the secretion of TNF- α and IL-10 from B cells. The statistical analysis was performed using one-way ANOVA with Tukey’s post-hoc test. Data are graphed as the mean ± SD (n = 4 tests, biological repeats). Source data are provided as a Source Data file.
Fig 4: MSC secretome enhancement via AIMS.a Schematic of the secretome enhancement via AIMS. b Heatmap presenting the secretome profiles of different groups after incubating for 4 days. The data was processed by the Z-score normalization and clustering analysis. c–j ELISA assay measuring the secretion of FGF-2, IGF-1, VEGF, HGF-1, IL-6, IL-10, IFN-γ, and TNF- α from different groups after incubating for 4 days. k The amounts of secreted exosomes from different groups measured by the EXOCET Exosome Quantitation Kit. l AchE activity of different groups after incubating for 4 days. m Secreted exosome numbers of different groups analyzed using NTA. n A representative TEM image of secreted exosomes from MSC aggregates. n = 3 tests. Scale bar: 100 nm. o NTA test measuring the diameters of secreted exosomes from different groups. The statistical analysis was performed using one-way ANOVA with Tukey’s post-hoc test. Data are graphed as the mean ± SD (n = 4, biological repeats). Source data are provided as a Source Data file.
Fig 5: The enhanced MSC secretome was attributed to increased cell-cell interactions mediated by N-cadherin.a Schematic illustrating how N-cadherin mediated cell-cell interactions contribute to the enhancement of the MSC secretome. b Heatmap measuring the secretome profiles of different groups after functional blocking of N-cadherin of MSCs and incubating for 4 days. The data was processed by the Z-score normalization and clustering analysis. c–j ELISA assay measuring the secretion of FGF-2, IGF-1, VEGF, HGF-1, IL-6, IL-10, IFN-γ, and TNF-α from different groups after functional blocking of N-cadherin of MSCs and incubating for 4 days (n = 4 tests, biological repeats). k Representative FDA staining images of different groups after incubating for 0, 12, 24 h, and 3 days, respectively. n = 3 tests with similar results. Scale bar: 100 μm. l Western blotting detecting the expression of N-cadherin of MSCs from different groups. n = 3 tests. a: control, b: 3D Microplate, c: 10 min, d: 30 min, e: 60 min. m Flow cytometry analysis measuring the expression of N-cadherin of MSCs from different groups. The statistical analysis was performed using one-way ANOVA with Tukey’s post-hoc test. Data are graphed as the mean ± SD. Source data are provided as a Source Data file.
Supplier Page from Abcam for Human Insulin like Growth Factor 1 ELISA Kit (IGF1)