Fig 2: Immunoexpressions of HOXC6 and Ki-67 in normal nasopharyngeal tissue (A,B); a high-stage, nasopharyngeal carcinoma (C,D), and a low-stage nasopharyngeal carcinoma (E, F), respectively.

Fig 3: HKDC1 expression potentiates cell proliferation, while HKDC1 knockdown reverses this effect in MCF7 cells. The MCF7 cells were infected by either expression or knockdown lentivirus for either PGC1ß or HKDC1, and the subsequent stable cell lines or related empty vector control (CTL) were cultured for 2 days, and then the cells were harvested for further analysis. (A) Cell proliferation analysis by thymidine incorporation, n = 5. (B) Colony formation assay in soft agar, n = 5. (C) Representative pictures for (B). (D) Quantitation of Ki-67 positive cells, n = 3. (E) Cell invasion and migration assay, n = 4. (F) Representative picture for (E). *, P < 0.05, vs. CTL group; ¶, P < 0.05, vs. ?PGC1ß group; #, P < 0.05, vs. shPGC1ß group. Results are expressed as mean ± SEM.

Fig 4: PHB2 knockdown by siRNA inhibits RD cell proliferation. (a) Evaluation of PHB2 knockdown by siRNA. RD cells were transfected with siRNAs against PHB2 (siPHB2-1 and siPHB2-2) or negative control (NC). Thirty-six hours later, cells were lysed and subjected to Western blotting using an anti-PHB2 antibody, and GAPDH was detected as a loading control. Full-length blots are presented in Supplementary Fig. S1. Results are representative of three independent experiments. (b) Effect of PHB2 knockdown on cell viability in RD cells. RD cells in 6-well plates were transfected with siRNAs. Thirty-six hours later, the cells were re-plated in triplicate in 96-well plates at 5000 cells per well. Cell viability was determined by the MTT assay. Data are presented as the mean ± S.D. (n = 3). Results are representative of three independent experiments. (c) Effect of PHB2 knockdown on expression of Ki-67. RD cells were fixed and stained 48 h after siRNA transfection and then monitored by immunofluorescence assay. Results are representative of three independent experiments. Scale bar 100 µm. (d) Effect of PHB2 knockdown on BrdU incorporation of RD cells. RD cells in 6-well plates were transfected with siRNAs. Twenty-four hours later, RD cells were re-plated in triplicate in 96-well plates at 5000 cells per well. Detection every 24 hours. Six hours before detection, RD cells were incubated with 10 µl/well of 10 µM BrdU labeling solution. Data are presented as the mean ± S.D. (n = 3). Results are representative of three independent experiments. (e) Effect of PHB2 knockdown on colony formation of RD cells. RD cells transfected with siRNA were plated in triplicate in 6-well plates at 1500 cells per well. After 10 days, images of the wells were taken. Data are presented as the mean ± S.D. (n = 3), *P < 0.05, **P < 0.01, ***P < 0.001, two-tailed Student’s t test. Results are representative of three independent experiments.

Fig 5: LMP1 knockdown in Epstein-Barr virus-positive tumor cells does not reduce PGC1ß-mediated tumorigenesis. SNK6 cells were treated with CTL, shLMP1, shNF-?B-p65 or shPGC1ß lentivirus before being harvested for biological assays. (A) mRNA levels were detected by quantitative PCR (n=4). (B) Representative western blot images. (C) Protein quantitation for panel B (n=5). (D) PGC1ß reporter activity (n=5). (E) Cell proliferation assay (n=5). (F) [3H]2-deoxyglucose uptake (n=5). (G) Colony formation assay (n=5). (H) Quantitation of Ki-67-positive cells (n=4). (I) Representative image of panel H. *P<0.05 vs. CTL group; ¶P<0.05 vs. shNF-?B-p65 group. LMP1, latent membrane protein 1; PGC1ß, peroxisome proliferator-activated receptor-? coactivator-1ß; sh, short hairpin; CTL, control; NF-?B, nuclear factor-?B.