Skip to main content
. 2021 Jun 8;12:3460. doi: 10.1038/s41467-021-23546-6

Fig. 5. Administration of VEGFC mRNA-LNPs results in non-significant blood vessel proliferation and immune response activation compared to the Poly(C) RNA-LNP control.

Fig. 5

a 1 µg of Poly(C) or VEGFC mRNA-LNPs was intradermally injected into the ear of C57Bl/6 mice and the growth of blood and lymphatic vessels were examined 22 days after the injection. Representative images of anti-CD31, anti-LYVE1, anti-Podoplanin, and anti-vWF stained paraffin-embedded ear samples are shown of 5 mice in each group (bars, 25 µm (anti-CD31, anti-LYVE1), 50 µm (anti-Podoplanin, anti-vWF)). Yellow arrows indicate LYVE1 positive lymphatic vessels, white arrows indicate CD31 positive and LYVE1 negative blood vessels. b The number of vWF high and LYVE1 negative blood vessels was determined 22 days after the administration of 1 µg of Poly(C) or VEGFC mRNA-LNPs. Data are represented as mean and SEM from slides of ears of 10 mice per group (two-tailed, paired T-test, P = 0.6344 for 10 mice). c Monitoring the effect on lymphatic endothelial cells and immune cells after intradermal injection of 1 µg of Poly(C) or VEGFC mRNA-LNPs into ears of Prox1GFP mice shown by flow cytometry analysis. Quantitative data for LYVE1+ , CD45+ , Ly6G/C (GR1)+, CD206+, CD3+, B220+ and CD11b+ Ly6G/C- cell numbers are represented as mean and SEM from Poly(C) or VEGFC mRNA-LNP-injected ears of 5–7 mice in each group, 22 days after mRNA-LNP injection (two-tailed, unpaired T-test, P = 0.0450 for LYVE1+ for 5 mice injected with Poly(C) and for 5 mice injected VEGFC mRNA-LNP, P = 0.5365 for CD45+ for 5 mice injected with Poly(C) and for 6 mice injected VEGFC mRNA-LNP, P = 0.1689 for Ly6G/C+ and CD45+ for 5 mice injected with Poly(C) and for 6 mice injected VEGFC mRNA-LNP, P = 0.7211 for CD206+ for 5 mice injected with Poly(C) and for 5 mice injected VEGFC mRNA-LNP, P = 0.8947 for CD3+ for 6 mice injected with Poly(C) and for 7 mice injected VEGFC mRNA-LNP, P = 0.7748 for B220+ for 6 mice injected with Poly(C) and for 7 mice injected VEGFC mRNA-LNP and P = 0.6922 for CD11b+ Ly6G/C- for 5 mice injected with Poly(C) and for 6 mice injected VEGFC mRNA-LNP comparing cell number). d Ly6G/C (GR1) positive immune cells were visualized after intradermal administration of 1 µg of Poly(C) or VEGFC mRNA-LNPs into the ears shown by anti-GR1 and anti-LYVE1 22 days after the injection. Representative images of anti-GR1 stained paraffin-embedded ear samples are shown of 3 mice per group (bars, 50 µm). Arrows indicate LYVE1 positive lymphatic vessels. Asterisks indicate P < 0.05 compared with control. All cell nuclei are labeled with DAPI (blue) in paraffin-embedded tissues.