Fig. 7: Genetic knockdown of KRT14 inhibits splenic metastasis.
From: EZH2-H3K27me3 mediated KRT14 upregulation promotes TNBC peritoneal metastasis
A, B The KRT14 knockdown was confirmed through immunoblot by using two different shRNA in 4T-1 and HCC1806 cells. The β-actin is used as a loading control. C–F Representative images of the wound healing assay to measure the migration ability of control and KRT14 KD in 4T-1 (C) and HCC1806 (E), magnification 10 ×. D, F The quantitative analysis of wound healing assay. Columns, a mean of quadruplicate readings of samples; error bar, ±SD, *P = 0.0001 and #P = 0.0001, compared to control, two-way ANOVA, Sidak’s multiple comparisons test. G–J Representative images of the trans-well chamber migration assay to measure the invasion ability of control and KRT14 KD 4T-1 (G) and HCC1806 (I) cells at 24 h. The quantitative bar graphs of the trans-well chamber migration assay of control and KRT14 KD 4T-1 (H) and HCC1806 (J) cells are shown. Columns represent a mean of triplicate readings of samples, error bars, ±S.D. compared to control, $P = 0.0002, %P = 0.0058, Student’s t-test (two-sided). K Fluorescence imaging of metastatic cells harvested from spleen (n = 5). The Td-Tomato fluorescence is for control cells, and GFP fluorescence is for KRT14 KD (left panel). Scale bar 50 μm. The excitation and emission wavelength: Td-Tomato-570–620 nm and GFP-465–520 nm. L The quantitative analysis for the metastatic control (Td-Tomato+) and KRT14 KD (GFP+) cells harvested from the spleen (n = 5, three different fields from each spleen), Columns, a mean of fifteen readings of samples, error bar, ±SD, &P = 0.0001, compared to Td-Tomato, Student’s t-test (two-sided). M Representative flow cytometry-derived scatter plots showing control (Td-Tomato+) and EZH2 Y641-F (GFP+) metastatic cells harvested from spleen (n = 5). N The quantitative analysis of the percentage of Control (Td-Tomato+) and KRT14 KD (GFP+) metastatic cells harvested from spleen (n = 5) Columns, a mean of five readings of samples, error bar, ±SD, @P = 0.0001, compared to Td-Tomato, Student’s t-test (two-sided). Source data are provided as a Source Data file.
