Fig. 3

TALAM1 synergizes with MALAT1 in breast cancer cell migration and invasion. a Quantification of MDA-MB-231 cells proliferative capacity up to 6 days after specific knockdown of MALAT1 or/and TALAM1 levels by determination of cellular metabolic activity through the resazurin-based assay (n = 3 ± SD. Two-sided Student’s t-test was used for statistical analysis; * p < 0.05; ** p < 0.01; *** p < 0.001). b Representative images of the wound healing assay performed in MDA-MB-231 cells after the specific knockdown of MALAT1 or/and TALAM1. c Percentage of wound closure quantified from the images acquired in the wound healing assay in (B) MDA-MB-231 cells (n = 3 ± SD. Two-sided Student’s t-test was used for statistical analysis; * p < 0.05; ** p < 0.01; *** p < 0.001). d Representative images of the wound healing assay performed in MCF10A cells exposed to 10 ng/ml of TGF-β after the specific knockdown of MALAT1 or/and TALAM1. e Percentage of wound closure quantified from the images acquired in the wound healing assay in (C) MDA-MB-231 cells (n = 3 ± SD. Two-sided Student’s t-test was used for statistical analysis; * p < 0.05; ** p < 0.01; *** p < 0.001). f Flow cytometry determination of CD44-positive cells from lung tissue (1 g) of mice injected with MDA-MB-231 cells downregulated for the MALAT1, TALAM1 or both. Cells treated with a control LNA were used as a control. (n = 3 ± SD. Two-sided Student’s t-test was used for statistical analysis; ** p < 0.01; *** p < 0.001) g) Percentage of mice presenting metastatic lesions after tail vein injection of MDA-MB-231 cells treated with the labelled conditions. Metastatic foci were assessed by an authorized pathologist and were composed of tumour cells with a solid pattern, with frequent mitosis. Number of mice per group is above the bars (Chi-squared test was used for statistical analysis; *** p < 0.001). h Representative microphotographs of the lung of mice injected with MDA-231 cells treated for 48 h with the labelled LNAs. Tumour metastasis (black arrow) was majorly seen with MDA-MB-231 cells treated with the control LNA. Metastatic foci (inside dashed line) expanded the lung (*, alveoli) and were composed of tumour cells with a solid pattern and high mitotic index (white arrowhead). Hematoxylin and Eosin. Scale bar 1 mm (upper panel) and 100um (lower panel)