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. 2023 Jan 4;6:9. doi: 10.1038/s42003-022-04392-8

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© The Author(s) 2023

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. 6 — a Representative images of micronuclei (white arrows) in mesenchymal cells isolated from the calvarias of Pfn1 knock-in (c.318_321del/WT) mice; scale bar 10 µm. b Quantification (%) of micronuclei in normal (WT/WT) and Profilin 1-deficient (c.318_321del/WT) mesenchymal cells; data are shown as mean ± s.e.m.; dots represent the value of each experiment (n = 4 biological replicates for WT samples, five biological replicates for mutant samples; scoring a total of 485 WT and 464 mutant cells; range of number of cells used for each individual experiment: 93-164 WT/WT, 67-107 c.318_321del/WT); *P = 0.0312. One-tailed unpaired Student’s t tests performed. c Quantification (%) of binucleation in normal (WT/WT) and Profilin 1-deficient (c.318_321del/WT) mesenchymal cells; data are shown as mean ± s.e.m.; dots represent the value of each experiment (n = 4 biological replicates for WT samples, five biological replicates for mutant samples; scoring a total of 646 WT and 711 mutant cells; range of number of cells used for each individual experiment: 103-234 WT/WT, 102-169 c.318_321del/WT); ns P = 0.0778. One-tailed unpaired Student’s t tests performed. d Images of primary Pfn1^WT/WT and Pfn1^{c.318_321del/WT} mesenchymal cells stained for F-actin (phalloidin, red) and nuclei (Hoechst 33342, blue); scale bars 50 µm. Arrowheads point towards trinucleated cells. Note that Profilin 1 insufficiency results in large cells with a loose actin cytoskeleton. e Representative haematoxylin/eosin-stained sections of the femoral epiphyseal growth plate of 4-month-old control (Pfn1^WT/WT) and knock-in (Pfn1^{c.318_321del/WT}) mice; scale bars 100 µm. Arrows point towards binucleated cells. f Quantification (%) of binucleated cells in the femoral growth plate of control (Pfn1^WT/WT) and knock-in (Pfn1^{c.318_321del/WT}) mice; data are shown as mean ± s.e.m.; dots represent the value of each experiment (n = 3 biological replicates; scoring ≥ 61 cells per replicate; **P = 0.0054. One-tailed unpaired Student’s t tests performed). g Haematoxylin/eosin-stained sections showing trinucleated cells (arrowheads) in 4-month-old Pfn1^{c.318_321del/WT} mice; scale bar 100 µm.