Fig 1: Ivermectin mediates mitochondrial dysfunction of ESCC cells. A. Intracellular ROS levels were investigated by DCFH-DA fluorescence in ESCC cells treatment with indicated; B. The expression levels of Bax and Bcl-2 were determined by western blot analysis; C. The expression of caspase cascade (cleaved-caspase 9, cleaved-caspase 3, PARP, cleaved PARP) were analyzed by western blotting; D. Mitochondrial membrane potential was observed by fluorescence microscope at 200 × magnifications, Scale bar = 50 μm; E. ATP production of ESCC cells after treated with ivermectin was detected; F. The mitochondrial DNA copy number in ESCC cells treatment with invermectin was evaluated using qRT-PCR. *P < 0.05, **P < 0.01.
Fig 2: Ivermectin suppressed xenograft growth of ESCC cells in vivo. A, B and C. Tumor growth curves and weight of subcutaneous xenograft tumor model developed from ESCC cells treatment with ivermectin as indicated (n=5); D and E. Representative immunohistochemistry images of Ki67, ROS1, p-p65 and cleaved-caspase 3 in xenograft tumor developed from KYSE-70 and KYSE-30 cells treatment with ivermectin as indicated. Scale bar: 300μm. *P < 0.05, **P < 0.01; ***P < 0.001.
Fig 3: Detection the levels of proteins in the section GBM xenograft tumor in four groups. (A) IHC was used to analyze the expression levels of PER2, G6PD and SIRT2 proteins in the section GBM xenograft tumor in four groups. (B) H&E staining and Bax, Bcl-2, Ros staining of tumor sections after RT in the four groups. (C) Immunofluorescence was performed to TUNEL in the section GBM xenograft tumor, and densitometric quantification of fluorescence ratio of TUNEL in the section GBM xenograft tumor. These results are expressed as the means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. Magnification, ×400. Ctrl: control, Met: metformin.
Fig 4: Metformin promoted apoptosis in GBM cell lines in vitro. (A) Cell cycle distribution detected by flow cytometry after metformin treatment at 24h, and quantified by the cell distribution rates. (B) Cell apoptosis detected by flow cytometry in metformin treatment and control groups at 24h, as quantified by the apoptosis rate. (C) JC-1 was detected by flow cytometry in metformin treatment and control groups at 24h, and quantified by the apoptosis rate. (D) ROS was detected by flow cytometry in the metformin treatment and control groups at 24h, and quantified by the apoptosis rate. Data are expressed as the means ± standard deviation (SD). *P < 0.05, **P < 0.01, ***P < 0.001. Ctrl: control, Met: metformin.
Supplier Page from Abcam for Anti-ROS1 antibody [EPMGHR2]