Fig. 2

CCL21 induced the up-regulation of MUC1 in ESCC cell lines. a, b the mRNA and protein expression level of MUC1 and CCR7 in ESCC cell lines; c: CCL21 induced the increasing MUC1 mRNA in ESCC cells. KYSE150, KYSE410, KYSE450 and Eca9706 were starved for 24 h and then treated with CCL21 at a concentration of 100 ng/ml for 12 h, then cells were harvested for the qRT-PCR; d, e CCL21 up regulated mRNA of MUC1 in a time and dose dependent way. Eca9706 and KYSE410 were starved for 24 h and then treated with CCL21 at a concentration of 0, 25, 50, 100, 200 ng/ml for 12 h or cells were treated with CCL21 at a concentration of 100 ng/ml for 0, 2, 6, 12 or 24 h. Then cells were harvested for qRT-PCR; f, g increasing of MUC1-C in KYSE410 and Eca9706 after treated with CCL21 at concentration of 0, 25,50, 100, 200 ng/ml for 24 h confirmed by immunoblotting; h, f blocking CCR7suppressed the up-regulation of MUC1 induced by CCL21. Eca9706 and KYSE410 were pretreated by CCR7 antibody (1 ug/mL) or IgG as control, then treated by 100 ng/ml for 24 h, then cells were harvested for detection of MUC1 by immunoblotting; j expression of MUC1 in KYSE410 and Eca9706 treated with PBS or CCL21(100 ng/mL) detected by immunofluorescence. Each data point represents the mean ± SD of three repeated experiments. *P < 0.05