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. 2021 Dec 31;33(6):694–707. doi: 10.21147/j.issn.1000-9604.2021.06.06

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Enrichment of TICs by heat intervention in vitro. (A,B) Western blot (A) and immunofluorescence analysis (B) of HSP70 expression by heat treatment of HepG2 cells. Nuclei were stained with DAPI; (C) Wound healing analysis for the closure ability of HepG2 cells after heat treatment; (D) Sphere-forming ability of HepG2 and Huh7 cells after heat treatment. The number of spheres more than 50 μm was quantified; (E) Proportions of CD133⁺ and EpCAM⁺ subpopulation of HepG2 cells in the indicated treatment groups were analyzed by FCA; (F) Western blot analysis for the stem cell-related genes expression of HepG2 cells in control and heat treatment group. Bar graphs in (B,D) are presented as . TIC, tumor-initiating cell; HSP, heat shock protein; DAPI, 4’,6-diamidino-2-phenylindole; EpCAM, epithelial cell adhesion molecule; FCA, flow cytometric analysis; SSC-A, side scatter area. ^**, P<0.01;^***, P<0.001.

Inline graphic — Enrichment of TICs by heat intervention in vitro. (A,B) Western blot (A) and immunofluorescence analysis (B) of HSP70 expression by heat treatment of HepG2 cells. Nuclei were stained with DAPI; (C) Wound healing analysis for the closure ability of HepG2 cells after heat treatment; (D) Sphere-forming ability of HepG2 and Huh7 cells after heat treatment. The number of spheres more than 50 μm was quantified; (E) Proportions of CD133⁺ and EpCAM⁺ subpopulation of HepG2 cells in the indicated treatment groups were analyzed by FCA; (F) Western blot analysis for the stem cell-related genes expression of HepG2 cells in control and heat treatment group. Bar graphs in (B,D) are presented as . TIC, tumor-initiating cell; HSP, heat shock protein; DAPI, 4’,6-diamidino-2-phenylindole; EpCAM, epithelial cell adhesion molecule; FCA, flow cytometric analysis; SSC-A, side scatter area. ^**, P<0.01;^***, P<0.001.