The effects of Jolkinolide B on the β-catenin pathway in HCC cells.

(A) Huh-7 and (B) SK-Hep-1 cells were treated with the indicated concentrations of Jolkinolide B for 48 h. The β-catenin protein expression was analyzed using western blotting. GAPDH was used as an internal control. The relative protein intensities were analyzed. (C, D) Huh-7 and SK-Hep-1 cells were treated with 10 μM Jolkinolide B for 48 h. The mRNA expression of MMP-7 and C-MYC in Huh-7 (C) and SK-Hep-1 (D) cells analyzed using RT-qPCR method. GAPDH was used as an internal control. *p p p p <0.0001.</p

MSI2 overexpression promotes β-catenin expression in HCC cells.

Huh-7 and SK-Hep-1 cells were transfected with lentivirus-mediated MSI2 overexpression plasmids for 48 h, the MSI2 protein expression (A) and MSI2 mRNA expression (B) were analyzed using western blotting and RT-qPCR methods, respectively. (C) β-catenin protein was detected in Huh-7 and SK-Hep-1 cells after MSI2 overexpression. The relative protein intensities were analyzed. *p p p <0.001.</p

Jolkinolide B downregulates the protein of MSI2 and the p53 expression in HCC cells.

(A, C) Huh-7 and (B, D) SK-Hep-1 cells were treated with the indicated concentrations of Jolkinolide B for 48 h. The MSI2 and p53 protein expressions were analyzed using western blotting. GAPDH was used as an internal control. *p p p <0.001.</p

S1 File -

Slide 1: Original images for Fig 2A, 2B, 2E. Slide 2: Original immunoblots for Fig 2C, 2D. Slide 3: Original immunoblots for Fig 3. Slide 4: Original immunoblots for Fig 4. Slide 5: Original immunoblots for Fig 5A, 5C. Slide 6: Original immunoblots for Fig 6A, 6B. Slide 7: Original immunoblots for Fig 6C. Sheet 1: Original CCK8 of cell viability for Fig 1A-1C. Original RT-qPCR for Figs 3C, 3D, 5B. (ZIP)</p

Jolkinolide B inhibits HCC cell lines migration, invasion and promotes HCC cell apoptosis.

(A) Huh-7 and SK-Hep-1 cells were seeded, scratched, and then treated with Jolkinolide B at a concentration of 10 μM for 48 h. The wound healing assays were performed to assess the migration abilities of Huh-7 and SK-Hep-1 cells. (B) Huh-7 and SK-Hep-1 cells were seeded in the upper transwell chamber and treated with DMSO or Jolkinolide B of 10 μM for 48 h to evaluate the migration and invasion abilities of HCC cells. (C) Huh-7 and SK-Hep-1 cells were treated with DMSO or Jolkinolide B at a concentration of 10 μM for 48 h; then the protein expression of E-cadherin and vimentin was analyzed using western blotting. GAPDH was used as an internal control. (D) Western blotting was used to assess Bax and BCL-2 protein expressions. GAPDH was used as an internal control. The relative protein intensities were analyzed. (E) Huh-7 and SK-Hep-1 cells were treated with DMSO or Jolkinolide B, the cells apoptosis was analyzed by flow cytometry and apoptosis rates were calculated. *p p p <0.001.</p

Jolkinolide B inhibits HCC cells proliferation.

Cell viability assays of Huh-7 (A), SK-Hep-1 (B) and L-02 cells (C) were performed using CCK-8 method after treatment with different concentrations (0, 5, 10, 25, 50, or 100 μM) of Jolkinolide B for 48 h.</p

Overexpression of MSI2 reverses Jolkinolide B-promoted apoptosis and Jolkinolide B-inhibited EMT and β-catenin signaling in HCC cells.

Huh-7 and SK-Hep-1 cells were treated with DMSO, 10 μM Jolkinolide B, or Jolkinolide B together with MSI2 plasmids. Then, the protein expressions of (A) Bax, BCL-2, (B) E-cadherin, vimentin and (C) β-catenin were analyzed using western blotting. GAPDH was used as an internal control. The relative protein intensities were analyzed. (D) Proposed working model of Jolkinolide B-induced inhibition of HCC. *p p p <0.001.</p