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Caption : stem markers increase in cells cultured with caf-cm and il6.
(a,b) quantitative real-time pcr (qrt-pcr) measuring csc-related genes cd44,cd24,aldha1,cd133,nanog,sox2,abcg2 and oct4 mrna levels in sk-br3 (a) and bt474 (b) cells treated with control, caf23-cm or naf1-cm for 24 h. data were normalized to gapdh mrna levels and expressed as fold change compared with controls (log2 scale). data are shown as mean ± se of 3 independent replicates for each condition. 
(c,d) quantitative real-time pcr (qrt-pcr) measuring csc-related genes cd44,cd24,aldha1,cd133,nanog,sox2,abcg2 and oct4 mrna levels in sk-br3 (c) and bt474 (d) cells treated with pbs or il6 for 24 h. data were normalized to gapdh mrna levels and expressed as fold change compared with controls (log2 scale). data are shown as mean ± se of 3 independent replicates for each condition. 


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Caption : apoptosis status in cells cultured with caf-cm and treated with trastuzumab alone or combined with multiple pathway inhibitors.
(a,b) apoptosis status in sk-br3 (a) and bt474 (b) cells were analyzed by staining with annexin-v/pi. cells were cultured with control medium or cm from caf64 and treated with trastuzumab (10ug/ml), or trastuzumab (10ug/ml) combined with il-6 neutralizing antibody (100 ng/ml), stat3 inhibitor s3i-201 (10 µm), bez235 (1 µm) or bkm120 (1 µm), nf-κb inhibitor bortezomib (10 nm), mek inhibitor u0126 (1 µm) for 24 h. the lower right quadrant (annexin-v+/pi−) and upper right quadrant (annexin-v+/pi+) indicate the percentage of early apoptosis and late apoptosis.


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Caption :  cell cycle in cells cultured with caf-cm and treated with trastuzumab alone or combined with multiple pathway inhibitors.
(a,b) cell cycles in sk-br3 (a) and bt474 (b) cells were analyzed by flow cytometry. cells were cultured with control medium or cm from caf64 and treated with trastuzumab (10ug/ml), or trastuzumab (10ug/ml) combined with il-6 neutralizing antibody (100 ng/ml), stat3 inhibitor s3i-201 (10 µm), bez235 (1 µm) or bkm120 (1 µm), nf-κb inhibitor bortezomib (10 nm), mek inhibitor u0126 (1 µm) for 24 h. these results were from one representative experiment of three independent trials.