Fig. 4: Expression of CKS2 contributes to cancer-associated phenotypes.

A, B Lentivirus knockdown of CKS2 in Y79 cells evaluated by qRT-PCR (A) and western blotting (B). GAPDH served as the internal control. C Proliferation rate evaluation of Y79 without (Scramble and Ctrl) and with CKS2 knockdown (CKS2_KD1 and CKS2_KD2) by CCK-8 assay. D Y79 cells were seeded in six-well plates at 1500 cells per well for colony formation assay. E Left panel: Representative images of EdU staining assay of Y79 cells without (Scramble and Ctrl) and with CKS2 knockdown (CKS2_KD1 and CKS2_KD2). Right panel: Quantification of percent EdU⁺ cells, shown as mean ± SEM, **p < 0.01, ***p < 0.001, (n ≥ 3). F, G Tumor xenografts using Y79 cells in CKS2 knockdown groups (CKS2_KD1 and CKS2_KD2) were smaller than without CKS2-KD groups (Scramble and Ctrl). Representative photographic images (F) and tumor weights (G) were shown (each group n = 4). H, I Lentivirus overexpression of CKS2 in CKS2-KD cells (CKS2_Res) restored expression of CKS2 in both RNA (H) and protein (I) levels. GAPDH served as the internal control. J Proliferation rate evaluation of Y79 cells between CKS2_KD group and CKS2_RES (Rescue) group by CCK-8 assay. K, L Colony formation assay (K) and EdU staining assay (L; Representative images at left panel and quantification of percent EdU⁺ cells at right panel^, shown as mean ± SEM, **p < 0.01, ***p < 0.001, n ≥ 3) of Y79 cells between CKS2_KD group and rescue group. M, N Tumor formation evaluation of Y79 cells between CKS2_KD group and CKS2_RES group. Representative photographic images (M) and tumor weights (N) were shown (each group n = 4). ***P < 0.001; **P < 0.01; *P < 0.05, by two-tailed t-test.