Fig. 5

LPS-enhanced HK3 expression in a P65 and Snail-dependent manner in CRC cells. a Established cells that stably knockdown Snail were treated with or without 1μg/ml LPS. The initial glucose concentration was 5mM in culture medium. Glucose consumption and lactate production were detected by glucose assay kit and lactic acid assay kit in the indicated times. b Scatter plot and Spearman correlation analysis of p65 and HK3 mRNA expression levels in TCGA database (left) and our tissue samples (right) of CRC respectively. Logarithmic transformed data were used in our tissue samples. ρ was Spearman’s rank correlation coefficient. c Scatter plot and Spearman correlation analysis of snail and HK3 mRNA expression levels in TCGA database (left) and our tissue samples (right) of CRC respectively. Logarithmic transformed data were used in our tissue samples. ρ was Spearman’s rank correlation coefficient. d The effects of P65 on HK3 expression were examined by western blotting. Cells were pretreated with 50 μM NF-κB inhibitor JSH-23 for 1 h, followed by 1μg/ml LPS treatment for 24 h. e Snail knockdown cells (shSnail) were stimulated with or without LPS to examine the effect of Snail on LPS-upregulated HK3. Protein expression was detected by western blotting. f Snail knockdown cells (shSnail) were transfected with HK3 overexpression vector under LPS treatment. Transwell assay was used to detect cell migration and invasion. Quantification of OD value was measured with microplate reader at 570 nm. Data are presented as mean±SEM. *P<0.05, **P<0.01, ***P<0.001. N, nuclear; p-P65, phosphorylated P65