Journal: Cell Death & Disease

Article Title: LGR5 promotes cancer stem cell traits and chemoresistance in cervical cancer

doi: 10.1038/cddis.2017.393

Figure Lengend Snippet: Overexpression and knockdown of LGR5 in human cervical cancer cell lines. ( a ) Immunohistochemical staining showing LGR5 expression in LGR5-overexpressing and LGR5-knockdown SiHa and HeLa cells, scale bar, 10 μ m. ( b ) A western blot assay was used to characterize the expression of LGR5 in LGR5-overexpressing and LGR5-knockdown SiHa and HeLa cells. ( c ) The expression of LGR5 in HeLa, SiHa cells was measured by western blot. ( d ) LGR5 expresssion was analyzed by flow cytometry. AcGFP: green fluorescent protein for control; LGR5: overexpression for LGR5; shLGR5: shRNA for LGR5; shControl: shRNA for control. Values are shown as the mean±S.D. * P <0.05; *** P <0.001

Article Snippet: Human cervical carcinoma cell lines HeLa and SiHa were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco Modified Eagle Medium (Sigma-Aldrich, St Louis, MO, USA), supplemented with 10% FBS (Invitrogen, Carlsbad, CA, USA), penicillin and streptomycin.

Techniques: Over Expression, Knockdown, Immunohistochemical staining, Staining, Expressing, Western Blot, Flow Cytometry, Control, shRNA

Journal: Cell Death & Disease

Article Title: LGR5 promotes cancer stem cell traits and chemoresistance in cervical cancer

doi: 10.1038/cddis.2017.393

Figure Lengend Snippet: LGR5-overexpressing cervical cancer cells exhibit enhanced self-renewal capacity. ( a ) Representative photos of tumorspheres formed by LGR5-overexpressing and LGR5-knockdown SiHa and HeLa cells. Bar, 50 μ m. ( b , c ) The number of tumorspheres/200 cells was counted from three consecutive passages. ( d ) The number of wells containing tumorspheres was counted. * P <0.05; ** P <0.01; *** P <0.001. Data represent mean±S.D. of triplicate experiments

Article Snippet: Human cervical carcinoma cell lines HeLa and SiHa were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco Modified Eagle Medium (Sigma-Aldrich, St Louis, MO, USA), supplemented with 10% FBS (Invitrogen, Carlsbad, CA, USA), penicillin and streptomycin.

Techniques: Knockdown

Journal: Cell Death & Disease

Article Title: LGR5 promotes cancer stem cell traits and chemoresistance in cervical cancer

doi: 10.1038/cddis.2017.393

Figure Lengend Snippet: Tumorigenic capacity of LGR5 LGR5+ and AcGFP LGR5 − cells in NOD/SCID mice from two cervical cancer cell lines

Article Snippet: Human cervical carcinoma cell lines HeLa and SiHa were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco Modified Eagle Medium (Sigma-Aldrich, St Louis, MO, USA), supplemented with 10% FBS (Invitrogen, Carlsbad, CA, USA), penicillin and streptomycin.

Techniques:

Journal: Cell Death & Disease

Article Title: LGR5 promotes cancer stem cell traits and chemoresistance in cervical cancer

doi: 10.1038/cddis.2017.393

Figure Lengend Snippet: LGR5 + cervical cancer cells are capable of differentiating in vitro and in vivo . ( a – d ) LGR5 + and LGR5 − cells were isolated from LGR5-overexpressing SiHa and HeLa or control cells and cultured in DMEM medium supplemented with 10% FBS and 1000 μ g/ml G418 for 2 weeks in vitro . The xenograft tumor cells from LGR5 + and LGR5 − cells in vivo were digested by collagenase IV overnight before detection. The expression of LGR5 was analyzed by flow cytometry

Article Snippet: Human cervical carcinoma cell lines HeLa and SiHa were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco Modified Eagle Medium (Sigma-Aldrich, St Louis, MO, USA), supplemented with 10% FBS (Invitrogen, Carlsbad, CA, USA), penicillin and streptomycin.

Techniques: In Vitro, In Vivo, Isolation, Control, Cell Culture, Expressing, Flow Cytometry

Journal: Cell Death & Disease

Article Title: LGR5 promotes cancer stem cell traits and chemoresistance in cervical cancer

doi: 10.1038/cddis.2017.393

Figure Lengend Snippet: Elevated LGR5 enhances migration, invasion and colony formation ability of cervical cancer cells in vitro. ( a , b ) The overexpression of LGR5 enhances migration in wound healing assays. Columns and error bars represent means±S.D. from one experiment of three independent experiments ( n =6 scratches per cell type and time point in each experiment). Scale bars, 800 μ m. ( c ) Cell migration and invasion was measured in Transwell chambers. Cells were counted with a microscope in nine random high-power fields. Migration and invasion ability was significantly increased in LGR5-overexpressing SiHa and HeLa cells. ( d ) Expression levels of EMT-related proteins in the LGR5-overexpressing and LGR5-knockdown HeLa and SiHa are demonstrated by western blotting. ( e , f ) Each 1000 cells were cultured in soft agar plates for 2 weeks. The number of colonies was assessed using crystal violet staining. Columns and error bars represent means±S.D. of three independent experiments using triplicate measurements in each experiment. * P <0.05; ** P <0.01; *** P <0.001

Article Snippet: Human cervical carcinoma cell lines HeLa and SiHa were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco Modified Eagle Medium (Sigma-Aldrich, St Louis, MO, USA), supplemented with 10% FBS (Invitrogen, Carlsbad, CA, USA), penicillin and streptomycin.

Techniques: Migration, In Vitro, Over Expression, Microscopy, Expressing, Knockdown, Western Blot, Cell Culture, Staining

Journal: Frontiers in Pharmacology

Article Title: Features and mechanisms of propofol-induced protein kinase C (PKC) translocation and activation in living cells

doi: 10.3389/fphar.2023.1284586

Figure Lengend Snippet: Sequential images of protein kinase C (PKC) fused with green fluorescent protein (PKC-GFP) translocation in HeLa cells elicited by various stimuli. Images were obtained using a confocal laser scanning microscope. (A) Application of 100 μM propofol elicited the translocation of PKCα-GFP to the plasma membrane (PM) within 1 min. PKCα-GFP was persistently localized in the PM during observation. Representative images of 28 observed cells are shown. (B) Application of 100 μM histamine elicited the translocation of PKCα-GFP to the PM within 30 s. PKCα-GFP almost completely returned to the cytoplasm within 5 min. Representative images of 12 observed cells are shown. (C) Application of 1 μM tetradecanoylphorbol 13-acetate (TPA) gradually and persistently translocated PKCα-GFP to the PM over 30-min of observation. Representative images of 12 observed cells are shown. (D) Propofol (100 μM) translocated PKCα-GFP to the nucleus in HeLa cells, in which intracellular calcium was eliminated by BAPTA-AM (10 μM). Representative images of 23 cells are shown. Bars indicate 10 μm.

Article Snippet: Materials were obtained from the following sources: propofol and histamine from FUJI FILM Wako Pure Chemical (Osaka, Japan), 12-O-tetradecanoylphorbol 13-acetate (TPA), 2,6-diisopropylphenol, and leptomycin B (LMB) from Sigma Aldrich (St. Louis, MO, United States), BAPTA-AM from Dojinido (Kumamoto, Japan), and glass-bottom culture dishes from MatTek Corporation (Ashland, OR, United States).

Techniques: Translocation Assay, Laser-Scanning Microscopy, Clinical Proteomics, Membrane