Figure 2.

CAIX is involved in the hypoxia-enhanced GBM migration and monocyte adhesion. (A) U87 and ALTS1C1 were treated with a CAIX inhibitor (U104; 3 μM) for 30 min and exposed to hypoxic conditions (1% oxygen (O₂)), and in vitro migration activities were measured after 24 h with a transwell assay and then visualized using a digital camera. (B) The quantification of U87 (upper panel) and ALTS1C1 (lower panel) migration by the number of cells that migrated to the underside of the filter. (C) U87 and ALTS1C1 were treated with a CAIX inhibitor (U104, 3 μM) for 30 min and exposed to hypoxic conditions (1% O₂) for 24 h. BCECF-AM-labeled-THP-1 was added to U87 and ALTS1C1 for 30 min, and subsequently, the adherence of THP-1 was measured by fluorescence microscopy. (D) The quantification of monocyte adhesion abilities by THP-1 cell adhesion on U87 (upper panel) or ALTS1C1 (lower panel). * p < 0.05 compared with the normoxia group. # p < 0.05 compared with the hypoxia group. A two-way analysis of variance with a post-hoc Bonferroni test was used to examine the significance of the mean. (E) U87 and U251 cells were transfected with empty vector or wild-type CAIX for 24 h and exposed to hypoxia (1% O₂) for 24 h. In vitro migration activities (left panel) and monocyte adhesion abilities (right panel) were measured using a digital camera and fluorescence microscopy, respectively. (F) The quantification of GBM migration abilities (upper panel) and monocyte adhesion abilities (lower panel). * p < 0.05 compared with the empty vector group (Student’s t-test). Quantitative data are presented as the mean ± standard error (representative of n = 3).