Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie
Article Title: β-HB treatment reverses sorafenib resistance by shifting glycolysis-lactate metabolism in HCC.
doi: 10.1016/j.biopha.2023.115293
Figure Lengend Snippet: Fig. 5. β-HB treatment reversed sorafenib resistance and enhanced regorafenib sensitivity by inhibiting the B-raf/MAPK pathway in Huh7-SR and Sk-Hep-1-SR cells. (A) Huh7-SR and SK-Hep-1-SR cells were treated with sorafenib (0 and 10 μM) and β-HB (0, 2.5, and 5 mM) for 48 h, and cell viability was then analyzed using an MTT assay. (B) Huh7-SR and SK-Hep-1-SR cells were treated with regorafenib (0 and 5 μM) and β-HB (0, 2.5, 5, and 10 mM) for 48 h, and cell viability was then analyzed using an MTT assay. (C) Western blotting was conducted to analyze the protein expression of phosphorylated (p)- and total (t)-MAPK-related signaling cascades; α-tubulin was used as an internal control. (D) Western blot images from Fig. 4 C were quantified using ImageJ software. (E) Representative contour plots of apoptosis were detected by flow cytometry stained with Annexin V-FITC and PI. (F) Apoptosis rates were assessed in dose-dependent regorafenib with or without β-HB treatment in Huh7-SR and Sk-Hep-1-SR with Annexin V/PI staining. * p < 0.05; * * p < 0.01; * ** p < 0.001 vs. 0 mM β-HB cells. Data are presented as mean ± SD.
Article Snippet: The following antibodies were used in the experiments: HMGCS2 (1:1000, #ab137043, Abcam, Cambridge, UK), hexokinase I (1:1000, #2024, Cell Signaling, Danvers, MA, USA), hexokinase II (1:1000, #2867, Cell Signaling), phosphofructokinase (1:1000, #8164, Cell Signaling), LDHA (1:1000, #3582, Cell Signaling), PDH (1:1000, #3205, Cell Signaling), IDH (1:1000, #8137, Cell Signaling), phosphorylated (p)- and total (t)-B-raf (1:1000, #2696 and #9433, Cell Signaling), p- and t-MAPK kinase (MEK; 1:1000, #2338 and #9122, Cell Signaling), ERK (1:1000, #9101 and #4695, Cell Signaling), β-catenin (1:1000, #8480, Cell Signaling), ZO-1 (1:1000, #8193, Cell Signaling), Vimentin (1:1000, #5741, Cell Signaling), N-cadherin (1:1000, #13116, Cell Signaling) and antiα-tubulin (1:5000 dilution, #T9026, Sigma-Aldrich).
Techniques: MTT Assay, Western Blot, Expressing, Control, Software, Flow Cytometry, Staining
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