Fig. 5

Secretion of CXCL12 is increased by ISO via β2AR-HIF-1α signaling in osteoblasts. qRT-PCR assay for metastasis related genes in MC3T3E1 (a) and primary osteoblasts (b) treated with 10 μM ISO for 12 h. The data are shown as fold changes compared to that in the control groups (n = 3). c CXCL12 level was detected by ELISA in MC3T3E1 and primary osteoblasts treated with 10 μM ISO for 24 h. (n = 4). d HIF-1α expression and quantification were determined by Western blotting in MC3T3E1 and primary osteoblasts treated with 10 μM ISO (n = 3). e HIF-1α expression and quantification were determined by Western blotting in MC3T3E1 and primary osteoblasts treated with 10 μM ISO combined with or without 50 μM ICI118,551 for 24 h (n = 3). f CXCL12 level was detected by ELISA in MC3T3E1 and primary osteoblasts treated with ISO combined with or without YC-1, a HIF-1α antagonist, for 24 h (n = 4). g MC3T3E1 and primary osteoblasts were transfected with siRNAs of HIF-1α or control siRNAs for 24 h and then stimulated with ISO for 24 h. Expression of HIF-1α were detected by Western blotting (left) (n = 2), and CXCL12 level was detected by ELISA (right) (n = 4). Data represent the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 and n.s. not significant