Figure 6. Long-term LC are homologous to eLC and up-regulate Id2.

A. Correlation matrix comparing differentially expressed genes between and blood monocytes, mLC 10 weeks post-BMT + T cells, and eLC from age-matched controls. B. Graphs show the relative FPKM count normalized to the maximum value for different transcription factors from the RNAseq data. Significance was calculated with a 1-way ANOVA, blood Ly6C⁺ monocytes n = 2, epidermal EpCAM⁺ cells n= 3, donor LC n = 3, age-matched eLC n = 3. C. BM cells were cultured for 6 days with GM-CSF, TFGβ and different combinations of BMP7, CSF1 and IL-34. Box and whiskers graph shows mean ± min. to max. numbers of DEC205⁺EpCAM⁺ cells in the cultures. Significance was calculated with a 1-way ANOVA for non-parametric samples with Dunn’s multiple comparisons test. Each symbol is data from one culture, n = 5 independent BM donors, in 3 independent experiments. D. The bar graph shows the mean expression ± SD of Runx3 or Id2 relative to GAPDH in sorted DEC205⁺EpCAM⁺ cells. Symbols are cells from 4 independent BM donors, in 3 independent experiments. Id2 expression in LC generated in the absence versus the presence of IL-34 was analyzed by paired t-test. E. Bar graph shows the mean frequency ± SD of EdU⁺ cells on day 6 of culture after cells where pulsed with EdU for 24 hours on day 2 or day 5. Symbols are cells from independent cultures (n = 2–4). Data was analyzed using a 1-way ANOVA. F. Line graph shows the frequency of viable DEC205⁺EpCAM⁺ LC in GM-CSF / TGFβ cultures with, or without, IL-34. Symbols represent paired individual BM cultures, and were analyzed using a paired t-test, n = 5. *P<0.05, **P<0.01, ***P<0.001.