Figure 3

RPLP1 is a candidate downstream gene in cervical cancer cells. (A) RNA –seq data from the analysis between SiHa cells with si-NC and si-CNN3#1 at 48 h post-transfection. Shown in the cluster map are differentially expressed genes (P. adjusted ≤0.05) in common (n = 48). Of those, 19 were up-regulated and 28 down-regulated. Data shown are from three independent experiments. The arrow indicates CNN3. (B) After literature review, 23 key candidate downstream molecules were selected and validated by RT-PCR in SiHa and CaSki cells treated with si-NC and si-CNN3#1, respectively. Each column represents the fold change of relative CNN3 mRNA expression (i.e., relative CNN3 mRNA expression of si-CNN3#1 group/that of si-NC group). Data shown are mean ± SEM (n = 3), *P < 0.05, ^#P < 0.01. (C) Immunoblot results of RPLP1 protein expression in cervical cancer cells with CNN3 upregulation and downregulation. At 48 h post-transfection, whole-cell lysates from SiHa and CaSki cells were collected to analyze the indicated proteins. β-actin was used as internal control. Quantification and statistical analysis of blots are shown (right graphs), mean ± SEM (n ≥ 3). *P < 0.05, **P < 0.01, ns indicates P > 0.05. Blots of CNN3 and β-actin cropped from same parts of the same gel, while blots of RPLP1 cropped from different parts of gel. The same is true for the blots in the following figures. Full-length blots are presented in Supplementary Fig. S5. (D) Representative immunoblot results of CNN3 protein expression from SiHa and CaSki cells with RPLP1 upregulation and downregulation. Quantification and statistical analysis of blots are shown (right graphs), mean ± SEM (n ≥ 3). *P < 0.05, **P < 0.01, ns indicates P > 0.05. Full-length blots are presented in Supplementary Fig. S6.