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. 2021 Apr 13;12:2198. doi: 10.1038/s41467-021-22522-4

Fig. 2. CRISPR/Cas9-mediated knock-out of the EFA6B encoding gene PSD4 in HMLE luminal and basal populations induces collective invasion in collagen I.

Fig. 2

a The cell surface marker EpCAM and CD49f were used to sort three epithelial cell populations including the luminal (light blue), luminal progenitors (orange) and mature basal cells (purple). These cells were immediately processed for CRISPR/Cas9-mediated PSD4 knock-out. b The HMLE WT population, the luminal progenitor clone WT23, heterozygous EFA6B KO25 (Het 25), homozygous EFA6B KO3, and the mature basal clone WT4, homozygous EFA6B KO1 cells were solubilized and the expression of the indicated proteins analyzed by immunoblot. Actin served as a loading control. c Representative images of the indicated cells grown 5 days in collagen I and stained for F-actin (red) and the nuclei (blue). Scale bars 20 μm. d Quantification of the percentage of cell aggregates (n = 100) with invasive protrusions of the indicated cell lines grown in collagen I for 5 days. N = 3 for WT, WT23, Het25, KO3; N = 5 for WT4 and KO1, average ±SEM, paired Student’s t-test p-values are versus WT23 for luminal cell lines and WT4 for basal cell lines. Source data are provided as a Source Data file.