Fig. 8: HK2 silencing in 4T1 cells inhibits metastasis to the lung, decreases SNAIL protein level and EMT gene expression.

a Immunoblot image, representative of 3 independent experiments, showing HK2 levels in two isogenic breast cancer cell lines: 67NR (a non-metastatic line) and 4T1 (a highly metastatic line). b Left: representative images showing H&E staining of lung sections after orthotopic transplantation of 4T1 or 4T1shHK2 cells into Balb/cj mice. Scale bar: 5 mm. Right: quantification of lung metastatic lesions (4T1 transplantation, n = 10; 4T1 shHK2 transplantation, n = 9) at endpoint. Results are the mean ± SEM; ***p = 0.0006 analyzed by an unpaired two-tailed t-test. c A representative immunoblot image showing the effect of HK2 silencing in 4T1 cells and overexpression of HK2 in 67NR cells on SNAIL and E-cadherin protein levels and GSK3β phosphorylation n = 3 (three independent experiments). d Quantification of protein levels after HK2 silencing in 4T1 cells. Results are the mean ± SEM of three independent experiments. *p < 0.05 versus 4T1 (HK2 p = 1.29E-06, HK1 p = 0.91, Vim p = 0.043, E-cad p = 0.0064, SNAIL p = 0.0033, P-GSK3B p = 0.0039, Total GSK3B p = 0.59); unpaired two-tailed t-test. e Quantitative RT-PCR analysis measuring relative mRNA levels in 4T1 and 4T1shHK2 cells. Results are the mean ± SEM (n = 4 biologically independent experiments). *p < 0.03 versus 4T1 (HK2 p = 5.7E-10, SNAIL p = 0.0023, E-cad p = 2.34E-06, Vim p = 0.026); unpaired two-tailed t-test. Uncropped blots are provided as a (Supplementary Fig. 16). Source data are provided as a Source Data file.