Fig. 5.

miR-23b overexpression and Btbd7 silencing both impair fSCS resistance in CRC cell lines and regulate in vivo cell invasiveness. (a) Left panel: representative images of fixed and stained post-fSCS colonies formed in 10 cm plates by HCT116 and HT29 after transfection of miR-23b precursor or scramble. Right panel: bars in the graph represent mean +/− std. of the number of colonies generated by scramble and miR23b overexpressing cells (ns, non-significant; **p < .005, using paired t-test with unequal variances) (b) Left panel: representative images of fixed and stained post-fSCS colonies formed in 10 cm plates by TN4_Sorted and TN4_20 cells after transfection of miR-23b precursor or scramble. Right panel: bars in the graph represent mean +/− std. of the number of colonies generated by TN4_Sorted and TN4_20 cells. (ns, non-significant; *p < .05, using paired t-test with unequal variances) (c) In vivo extravasation assay. Mice were intravenously injected with DiI labeled TN4_Sorted and TN4_20 cells overexpressing miR-23b precursor or scramble. After 72 h, infiltrated cells in the lung parenchyma were quantified as described in Material and Methods. Dot plot indicates mean +/− std. of the fraction of infiltrated cells over total nuclei in lungs of 3 mice per condition. (d) Left panel: representative images of fixed and stained post-fSCS colonies formed in 10 cm plates by TN4_Sorted and TN4_20 cells after silencing of BTBD7 with Sh-BTBD7. Right panel: bars in the graph represent mean +/− std. of the number of colonies generated by TN4_Sorted and TN4_20 cells (ns, non-significant; *p < .05, using paired t-test assuming unequal variances).