Fig. 2.

UCKL1 knockdown diminishes CRC cells growth. (a) Real-time qPCR results of UCKL1 mRNA level in doxycycline-inducible UCKL1 shRNA transduced HCT116 or RKO cells treated with or without doxycycline. Unpaired two-tailed Student’s t-test was used to compare the results. (b) Western blot results of UCKL1 protein level in doxycycline-inducible UCKL1 shRNA transduced HCT116 or RKO cells treated with or without doxycycline. (c) Cell viability of doxycycline-inducible UCKL1 shRNA transduced HCT116 or RKO cells treated with or without doxycycline. The data are represented as the means ± SD. Two-way ANOVA was used to compare the results. (d) Colony formation rates of doxycycline-inducible UCKL1 shRNA transduced HCT116 or RKO cells treated with or without doxycycline. Quantification of the colony formation rates are shown in right panel. The data are represented as the means ± SD. Unpaired two-tailed Student’s t-test was used to compare the results. (e) Wound healing results of doxycycline-inducible UCKL1 shRNA transduced HCT116 or RKO cells treated with or without doxycycline. Scale bars: 50 μm. Quantitative analyses are shown in right panel. The data are represented as the means ± SD. Unpaired two-tailed Student’s t-test was used to compare the results. (f) The efficacy of UCKL1 knockdown on tumour growth was examined by subcutaneous injection of RKO cells into nude mice. UCKL1 knockdown mice were fed with water containing doxycycline (2 mg/ml), while control mice were just fed with water. Tumour growth curves are shown in the left panel. Tumour volume = length × width² × 1/2. The data are represented as the means ± SD (n = 5). Two-way ANOVA was used for comparison of differences between growth curves. Representative images of tumours are shown in right panel.