Inhibition of nuclear translocation of p65 has no effect on the hypoxic induction of various genes in human MDMs in vitro. (A-B) Effect of the p65 inhibitor JSH-23 (or its vehicle, DMSO) on the nuclear translocation of p65 induced by TNF-α or hypoxia by human MDMs. N indicates normoxia; H, hypoxia (0.1% O₂); % nuclear p65 immunofluorescence, the percentage of the total, DAPI-stained (blue) area of MDM nuclei that was GFP⁺ (green). The figures at the base or just above each bar represent the average percentage of all MDM nuclei immunofluorescent for p65 (B). *P < .05 with respect to normoxia alone group. ⁺P < .05 with respect to TNF-α + DMSO group. ∧P < .05 with respect to hypoxia + DMSO group. (C) Effect of JSH-23 on NF-κB binding and accumulation of HIFs-1 and -2α in hypoxic MDMs. N indicates normoxia; H, hypoxia, H + JSH-23, hypoxia after JSH.23 treatment. All 3 groups received the vehicle for JSH-23, DMSO. A vertical line has been inserted to indicate repositioning of lanes from the same gel; (1) left panel: EMSA showing NF-κB binding to a DNA consensus sequence, and (2) right panel: immunoblots for HIFs-1 and -2α. (D) Effect of JSH-23 blockade of p65 function on the fold induction of VEGFA, CXCL8, IL-1β, CXCR4, GLUT-1, STAT4, ADM, and ADORA2A by TNF-α or hypoxia. *P < .05 with respect to normoxia with DMSO alone. **P < .05 with respect to group indicated. $P < .05 with respect to TNF + DMSO group. Pooled data from 3 replicate experiments are shown.