Fig 1: Knockdown of WNK1 suppressed the malignant behaviors of HCC cells by suppressing autophagy and inactivating AMPK pathway. (a, b) si-AMPK transfection reduced the expression patterns of AMPK, Beclin 1, LC3 II/I, and increased P62 level, and the above results were reversed after si-WNK1 treatment. (c, d) si-AMPK transfection increased the OD values of Huh7 and Hep3B cells, while si-WNK1 treatment counteracted the effect of si-AMPK transfection on the OD values of Huh7 and Hep3B cells. (e–g) si-AMPK transfection increased the clone number of Huh7 and Hep3B cells, while si-WNK1 treatment counteracted the effect of si-AMPK transfection on the clone number of Huh7 and Hep3B cells. ∗p < 0.05, ∗∗p < 0.01 vs. control, ##p < 0.01 vs. si-WNK1, &p < 0.05, &&p < 0.01 vs. si-AMPK.
Fig 2: Loss of WNK1 reduced cell cycle, invasion, and migration in HCC cells. (a, b) Data from GEPIA website showed the relationship between WNK1 and CDK1/CDK2. (c, d) Loss of WNK1 reduced the protein patterns of CDK1 and CDK2 in Huh7 and Hep3B cells. (e) Loss of WNK1 decreased the invasion number of Huh7 and Hep3B cells. (f) Loss of WNK1 diminished the migratory area of Huh7 and Hep3B cells. ∗∗p < 0.01 vs. si-con.
Fig 3: Loss of WNK1 promoted autophagy and AMPK pathway. (a, b) Loss of WNK1 increased the protein patterns of Beclin 1 and LC3 II, and reduced P62 level. (c, d) Immunofluorescence staining was used to detect LC3 II immunoreactivity. (e, f) Loss of WNK1 increased the protein patterns of p-AMPK in Huh7 and Hep3B cells. ∗∗p < 0.01 vs. si-con.
Fig 4: WNK1 was highly expressed in HCC cell lines and promoted cell growth. (a, b) WNK1 expression was significantly increased in HCC cell lines. (c, d) The treatment of si-WNK1 or si-WNK2 obviously reduced WNK1 expression. (e) The relationship between WNK1 and PCNA was analysed by GEPIA website. (f, g) Loss of WNK1 significantly reduced the OD values of Huh7 and Hep3B cells. (h–j) Loss of WNK1 reduced the number of clones in Huh7 and Hep3B cells. ∗∗p < 0.01 vs. si-con.
Fig 5: Loss of WNK1 restricted the malignant behaviors of HCC cells by suppressing autophagy and inactivating AMPK pathway. (a, b) GSK690693 treatment reduced the expression patterns of p-AMPK, Beclin 1, LC3 II/I, and increased P62 level, and the above results were reversed after si-WNK1 treatment. (c, d) GSK690693 treatment increased the OD values of Huh7 and Hep3B cells, while si-WNK1 treatment counteracted the effect of GSK690693 on the OD values of Huh7 and Hep3B cells. (e–g) GSK690693 treatment increased the clone number of Huh7 and Hep3B cells, while si-WNK1 treatment counteracted the effect of GSK690693 on the clone number of Huh7 and Hep3B cells. ∗p < 0.05, ∗∗p < 0.01 vs. control, ##p < 0.01 vs. si-WNK1, &p < 0.05, &&p < 0.01 vs. GSK690693.
Supplier Page from Abcam for Anti-WNK1 antibody