Fig. 3: FRMD3 decreased the mobility of BRCA cells and their growth and metastasis in vivo.

a GFP fluorescence microscopy (left) and Western blot (right) analysis of FRMD3 levels of T47D and MDA-MB-231 cell lines stably overexpressing GFP-FRMD3 established using lentivirus. Scale bar, 100 μm. b Cell tracking assay on three cells of MCF10 cells with FRMD3 knockdown or BRCA cells with GFP–FRMD3 overexpression, respectively. Images were captured every hour, and the trajectories of representative cells are plotted. The origins of migration are superimposed at (0, 0). c–f T47D (c, d) and MDA-MB-231 (e, f) cell lines stably overexpressing FRMD3-GFP, along with control stable cells, were injected subcutaneously into the mammary fat pads of female nude mice (n = 6 per group). Gross pictures (c, e), volumes (left in d, f) and weight (right in d, f) of xenografts were shown. Scale bar, 1 cm. g RT-qPCR (left), IHC (middle) and IF (right) analysis of the mRNA or protein levels of FRMD3 in the xenografts. Scale bar, 100 μm. h Representative in vivo GFP fluorescence images of primary and metastatic tumors derived from MDA-MB231 cells with FRMD3-GFP overexpression. Scale bar, 100 μm. i Representative images (left) of the lungs from BALB/c nude mice 30 days after tail vein injection with stable MDA-MB-231 cells with or without FRMD3-GFP overexpression and the number of metastatic nodules on the surface of the lungs from each group (right). (n = 6 per group). j Representative images of H&E staining of mouse lung tissues in Fig. 3i. Scale bars: 200 μm (left panel) and 50 μm (right panel). N = 3 biologically independent replicates. The student’s t-test was used to estimate the significance of difference between two groups. Data were presented as means ± s.d. ***P < 0.001.