Fig. 5.

Knockdown of lnc273–31 or lnc273–34 inhibits the self-renewal maintenance of colorectal CSC in vitro and in vivo. a. Migratory and invasive ability of HCT116 p53 endogenous point mutant (PM) cells with lnc273–31 (upper) or lnc273–34 (down) knockdown were measured by transwell migration and matrigel invasion assays using ASOs. ASO, antisense oligos. b. The spheroids size (left) and numbers (right) of 1st spheroid formation (upper) and 2nd spheroid formation (down) assay after lnc273–31 (upper) or lnc273–34 (down) silenced in HCT116 p53 endogenous point mutant (PM) cells. Scale bar, 500 μM. c. ALDH enzymatic activity was assessed using the Aldefluor assay system in lnc273–31 or lnc273–34 silencing cells. d. Stable lnc273–31/34-knockdown or shNC spheroid cells were diluted and subcutaneously implanted into BALB/c nude mice and tumor-free mice were monitored. N = 5 for each group. e. Stable lnc273–31/34-knockdown or shNC spheroid cells were subcutaneously injected into BALB/c nude mice for observation of tumor growth. The tumor volumes were measured at the indicated time points (left), and images of tumors from nude mice at autopsy are presented (right). f. Tumorigenic cell frequency in stable lnc273–31/34-knockdown or shNC spheroid cells was determined with ELDA analysis (http://bioinf.wehi.edu.au/software/elda/). CI, confidence interval. The experiments were performed in triplicate (a-c). Data are shown as means ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001