Fig. 4.

Motility of DCs differentiated under hypoxia is reduced. a Incubation time for the random migration assay was 16 h. The random migration ability of H/H-imDCs was decreased by 50%. The average of three independent experiments is shown. Bars show the means plus SD. **P < 0.01 versus N/N-DCs is shown. b Expression of CCR5 and CCR7 was analyzed by FACS analysis (left panel). CCR5 expression was found in imDCs but not in mDCs, and the expression intensity was very similar between N/N-, N/H-, and H/H-imDCs (upper panel). CCR7 expression was found in mDCs but not in imDCs except for H/H-imDCs (lower panel). Isotype and DCs are presented as light gray and thin black lines in the histogram. To evaluate the effects of the hypoxia on chemokine receptor function, the chemotactic responsiveness was assessed (right panel). The number of migrating H/H-imDCs toward CCL5 was decreased by 50%. Similarly, that of H/H-mDCs toward CCL21 was also decreased by 65%. Right panels the average of three independent experiments is shown. Bars show the means plus SD. **P < 0.01 versus N/N-DCs is shown. c DCs that invaded through the Matrigel-coated filter were examined and were counted under a light microscope (× 100) (upper panel). The number of invading H/H-imDCs was decreased by 78% (lower left panel). The number of invading H/H-mDCs was decreased by 73% when compared with N/N- or N/H-mDCs (lower right panel). The average of three independent experiments is shown. Bars show the means plus SD. **P < 0.01 versus N/N-DCs is shown. d The number of invading MMP9-silenced DCs was decreased