Fig. 3: CXCL12 is mainly produced by luminal epithelial cells and is required for mammary ductal morphogenesis. | Nature Communications

Fig. 3: CXCL12 is mainly produced by luminal epithelial cells and is required for mammary ductal morphogenesis.

From: CXCR4+ mammary gland macrophageal niche promotes tumor initiating cell activity and immune suppression during tumorigenesis

Fig. 3

a IF images of CXCL12 with Keratin 14 (K14) or Keratin 8 (K8) in WT mammary epithelium. b Representative flow cytometry plots showing CXCL12 protein and mRNA expression in basal/MaSCs and luminal cells. c, f Representative carmine alum-stained mammary glands from K8-CreER control (Ctrl) and CXCL12K8-cKO (cKO) mice at 6 and 14 weeks. d, e, and g Quantification of ductal length (d), and branching (e and g) in week 6 (n = 10, 14) and week 14 (n = 4, 6, biologically independent samples) mammary glands. h Quantification of Ki67+ epithelial cell in control vs. CXCL12K8-cKO mammary glands (n = 4, 5, biologically independent samples). i F4/80 and K8 IF staining of control and CXCL12K8-cKO mammary glands. Right panels show enlarged boxed regions. j Quantification of intraepithelial F4/80+ cell per field (n = 3, biologically independent samples, 2–3 ductal areas were randomly selected per each gland section for quantification). k Flow cytometry quantification of Lin−CD24+CD29Hi P4 cells (basal/MaSCs) (Supplementary Fig. 16a) in control and CXCL12K8-cKO mammary glands (n = 9, 11, biologically independent samples). l Mammosphere formation assay with 5000 P4 cell from control and CXCL12K8-cKO (n = 3, biologically independent samples). m, n Limited dilution assay with basal/MaSCs from control and CXCL12K8-cKO mammary glands. Representative mammary outgrowths. Table representing serial dilution injections with the corresponding take rate and repopulation frequencies (calculated by ELDA, Pearson’s Chi-squared test, two-sided). Scale bar, 50 μm (a and i), 25 μm (c, f, and m). Data are shown as mean values  ±  s.d. Statistical significance was calculated by two-tailed unpaired Student’s t-test or one-way ANOVA with Turkey’s test. Box plots show the median (center line), with box bounds representing the 25th and 75th percentiles (lower and upper quartiles). Whiskers extend to the most extreme values within 1.5x the interquartile range (IQR); points beyond this range are plotted as outliers. Source data are provided as a Source Data file.

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