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. Author manuscript; available in PMC: 2021 Jan 12.
Published in final edited form as: Nature. 2019 Jan 16;565(7740):505–510. doi: 10.1038/s41586-018-0858-8

Extended Data Figure 5. Mono-cultures of vascular cells in diabetic media.

Extended Data Figure 5

a, iPSC (NC8) derived endothelial cells cultured in non-diabetic conditions, in 75mM D-glucose (hyperglycemia), 1ng/mL TNFα+ 1ng/mL IL6, or diabetic media (hyperglycemia + TNFα+IL6), were stained for CD31 and Col IV. Right panel shows quantification of Col IV intensity of individual cells endothelial cells. The data is presented as mean ± S.D. from n=3 independent experiments. p values are indicated in the panel (One-way ANOVA) b, iPSC (NC8) derived vascular smooth muscle cells (vSMCs) were cultured in the presence of normal medium, hyperglycemia, TNFα+IL6 or diabetic media (hyperglycemia + TNFα+IL6). Cultures were stained for SMA, Col IV and Fibronectin (FN). Col IV and FN intensity of individual cells was quantified. The graph represents mean ± S.D. from n=3 independent experiments. P-values are indicated in the panel (One-way ANOVA) c, G1S1 endothelial cells were cultured in non-diabetic normal media and diabetic media (hyperglycemia + TNFα+IL6) and stained for Fibronectin (FN), Laminin or Collagen IV (Col IV). Phalloidin was used to visualize the actin cytoskeleton. Experiments were repeated independently n = 3 times with similar results. d, Col4a1expression was measured by RT-qPCR from G1S1 cells cultured in non-diabetic and diabetic media for different time points, as indicated. Expression was normalized to HPRT expression. n=3 independent experiments. P-values are indicated in the panel (Two-tailed t-test) Scale bars a,b,c=100μm.