Fig. 3: Rank deletion in luminal cells results in parity-associated protein translation defects.
From: Luminal Rank loss decreases cell fitness leading to basal cell bipotency in parous mammary glands
a Experimental protocol used to characterize the difference between virgin and parous K8imTmG and K8iΔRank females (for details see methods). b Quantification of GFP+ within K8+ cells in age-matched virgin (n = 3 K8imTmG; n = 4 K8iΔRank; p = 0.98) and parous MGs (n = 3 K8imTmG; n = 5 K8iΔRank; p = 0.93). c Percentage of luminal differentiated cells (PR+) in the K8+ lineage in virgin (n = 3 K8imTmG; n = 4 K8iΔRank; p = 0.11) and parous stage (n = 3 K8imTmG; n = 5 K8iΔRank; p = 0.01). d Quantification of proliferation (Ki-67) in luminal (K8+) cells in virgin (n = 3 K8imTmG; n = 4 K8iΔRank; p = 0.67) and parous (n = 3 K8imTmG; n = 5 K8iΔRank; p < 0.0001) MGs. e GSEA profiles of alveolar progenitor differential gene set obtained at G14.5 MG (AVD) between virgin and parous luminal (Lu) cells of the indicated genotypes. f qPCR analysis of differentially expressed lactation genes (Csn2 (p = 0.33), Csn3 (p = 0.006), Lalba (p = 0.06), Irx1 (p = 0.005), Btn1a1 (p = 0.03)) between virgin K8imTmG (n = 4) and K8iΔRank (n = 3) mice in sorted luminal GFP+ cells. g Bubble plot representation showing selected significant gene sets related to ribosomal/translation in the different biological settings indicated from luminal cells of K8imTmG and K8iΔRank mice. h Representative images of protein synthesis signal (OPP signal in magenta) in primary luminal cells (GFP+ in green) and quantification of protein synthesis (average intensity of OPP signal per cell/per field) from virgin (p = 0.03) and parous (p = 0.01) K8imTmG and K8iΔRank mice (n = 3). Data are represented as mean +/− SEM. Scale bars and significant P values are indicated in the graphs. P-values were calculated by Two-Way ANOVA with Tukey’s multiple comparisons (b, c, d, h) and One Sample T-Test (f). Staining was quantified in 5 independent images from two tissue sections collected 100 µm apart (b, c, d). Staining was quantified in 5 independent images (h). Source data are provided as a Source Data file. ns not significant.
