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. 2020 Dec 8;12(12):3679. doi: 10.3390/cancers12123679

Figure 2.

Figure 2

Effects of CWS-loaded formulation on growth inhibition in bladder cancer cells. (A) Human bladder cancer 5637 and HT1376 cells, and murine bladder cancer MBT2 cells were seeded in 96-well plates. Cells were treated with indicated concentrations of CWS-loaded formulations for 48 h, and then, cell viability was determined using MTT solution. * p < 0.005, ** p < 0.0005, ns: non-significant; untreated/CWS, untreated/CWS-Nano, or untreated/CWS-Nano-CL. Data are mean ± SEM (n = 6). (B) Colony-forming ability of cells treated with CWS-loaded formulations (1, 5, and 10 µg/mL) was measured after 2 weeks. * p < 0.005, ** p < 0.0001; untreated/CWS, untreated/CWS-Nano, or untreated/CWS-Nano-CL. Data are mean ± SEM (n = 3). (C) Cell numbers were counted by staining with trypan blue at 24, 48, and 72 h after treatment with CWS-loaded formulation. * p < 0.05, ** p < 0.0005; untreated/CWS or untreated/CWS-Nano-CL. Data are mean ± SEM (n = 3). (D) Levels of cleaved poly(ADP-ribose) polymerase (PARP) in cells treated with CWS-loaded formulations were analyzed by Western blotting. Actin was used as a loading control. The blots are representative of three independent experiments. The quantification graphs represent cleaved PARP (Cl’vd PARP)/Actin ratios determined by densitometric analyses. All expression ratios were normalized to the untreated group.