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. 2021 Oct 13;12:5981. doi: 10.1038/s41467-021-26269-w

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — A Peritumoral injection. Left: Image of a mouse with B16 melanoma after peritumoral injection of Evans Blue, injection site indicated by the arrow, fur removed for better visibility; right: Confocal microscopic image of the tumor margin after peritumoral injection of Oregon Green (OG)-labeled micelles. B Frequency of OG-labeled cells in different organs 1 and 24 h after peritumoral injection of OG-labeled L-PM micelles at day 14 B16-OVA melanomas. Representative flow cytometric plots, numbers represent cell frequencies in gates. LN lymph nodes (n = 6 (1 h) and n = 3 (24 h) per organ, all comparisons ***P < 0.001 (parametric one-way ANOVA, with Holm–Šídák’s multiple comparisons test, alpha = 0.05, 0.95 confidence interval). C Micelles (NL-PM, blue; L-PM, red), free inhibitor (MDL, gray), and solvent (DMSO, white) toxicity in tumor tissue 1 and 4 h after peritumoral injection at day 14 melanomas. Violin plots show median, interquartile range, minimum and maximum (truncated above and below max and min values) of dead CD45^negCD29⁺ tumor cells, n = 5 per treatment, ***P = 0.00099 (1 h) ***P = 0.00099 (4 h) ns = nonsignificant (parametric One-way ANOVA, with Holm–Šídák’s multiple comparisons test, alpha = 0.05, 0.95 confidence interval). D Intracellular cAMP levels in day 14 B16-OVA melanoma tissue after peritumoral injection of 10 µg MDL dissolved in DMSO, L-PM (10 µg MDL) or NL-PM (same amount of particle substance as L-PM). Data shown as mean ± SEM, n = 3 per treatment, *P = 0.006 (NL-PM vs. L-PM, 4 h), ***P = 0.00099 (NL-PM vs. L-PM, 1 h), ***P = 0.00099 (MDL vs. NL-PM, 1 h) (parametric one-way ANOVA, with Holm–Šídák’s multiple comparisons test, alpha = 0.05, 0.95 confidence interval). All figures and graphs are representative of three independent experiments each.