PMC

Inhibition of p600 does not affect cell proliferation, apoptosis, and colony-formation capacity under anchorage-dependent conditions. a Proliferation assay under anchorage-dependent conditions. There was no significant difference between p600-targeting siRNA-transfected MKN45 cells and control siRNA-transfected MKN45 cells in terms of cell proliferation under anchorage-dependent conditions. b Proportion of apoptotic cells in (i) control siRNA-transfected MKN45 cells and (ii) p600-targeting siRNA-transfected MKN45 cells as analyzed by flow cytometry. There was no significant difference between p600-targeting siRNA-transfected MKN45 cells and control siRNA-transfected MKN45 cells in terms of apoptotic cells proportion. c Macroscopic appearance of colonies formed under anchorage-dependent conditions; (i) control siRNA-transfected MKN45 cells and (ii) p600-targeting siRNA-transfected MKN45 cells. (iii) Data graph. Data are expressed as mean ± SD. There was no significant difference between p600-targeting siRNA-transfected MKN45 cells and control siRNA-transfected MKN45 cells in terms of colony-formation capacity under anchorage-dependent conditions

Pictures of different cell lines at a magnification of 10X
A - anchorage-dependent HeLa
B - anchorage-dependent with enhanced expression of siat7e
C - anchorage-dependent with reduced expression of lama4
D - anchorage-independent HeLa
E - anchorage-independent with reduced expression of siat7e
F - anchorage-independent with enhanced expression of lama4

Cell lysates prepared from (a) cell line derived tumour xenografts or (b) anchorage-dependent (2D) cell culture were immunoblotted using the indicated antibodies. (c) Expression levels of pChk1 (S296) and total Chk1 were determined by densitometry from (a,b) above.

Treatment of anchorage-dependent and anchorage-independent hepatoma cells with apyrase and eATP. a Anchorage-dependent and anchorage-independent BEL7402 cells were treated with 2.5 mM of eATP in the absence or presence of apyrase (1 U/mL). Cell viabilities were measured by CCK-8 assay after incubation for 24 h. b Anchorage-dependent and anchorage-independent BEL7402 cells were pretreated with apyrase for 0.5 h, followed by 2.5 mM of eATP treatment for 24 h before harvested for Western blot analysis. c–l Caspase 3 (c, d), LC3-II (e, f), p-AMPKα (g, h), p-mTOR (i, j), and p-S6K1 (k, l) in two types of cells were quantified by densitometric analysis and normalized to β-actin (c, e, g, i, and k for anchorage-dependent model and d, f, h, j, and l for anchorage-independent model). Presented figures are representative data from three independent experiments. *P < 0.05; **P < 0.01