Fig. 3

Knockdown of UCHL3 inhibited cell growth, colony formation, tumor formation, and tumor stem-like properties. a The MTS assay was used to assess cell viability in H358 cells with stable UCHL3 knockdown (n = 5). Data are shown as the mean ± SD; ****p < 0.0001. b A colony formation assay in plates was performed to detect the colony formation ability of H358 cells with stable UCHL3 knockdown (n = 3); representative images are shown in the Supplementary section, and the results show that knockdown of UCHL3 inhibited colony formation. Data are shown as the mean ± SD; **p < 0.01. c The H358 cell line with UCHL3 knockdown was seeded in ultralow attachment dishes to allow tumor sphere formation, and the results are shown as a bar graph (n = 3, scale bar = 100 μm). Data are shown as the mean ± SD; **p < 0.01. d Representative images from flow cytometry analysis to detect CD338-positive cells among UCHL3-knockdown H358 cells, with the results shown as a bar graph (n = 3). Data are shown as the mean ± SD; **p < 0.01. e Flow cytometry analysis of ALDH activity in the UCHL3-knockdown H358 cell line, with the results shown as a bar graph (n = 3). Data are shown as the mean ± SD; ****p < 0.0001. f–h A xenograft model of tumor growth was established to evaluate the ability of H358 cells with stable UCHL3 knockdown to form tumors. Tumor formation was monitored at the indicated times (f), and images (h) and tumor weights (g) were recorded (n = 6 mice per group). Data are shown as the mean ± SD; **p < 0.01, ****p < 0.0001. Data in all bar graphs were assessed by one-way ANOVA with multiple comparisons