Figure 6.

Clotrimazole induced dendritic cell activation by regulating Chop expression. (A and B) DC2.4 cells were treated with clotrimazole or DMSO for 24 hours, and the expression of Chop was detected by WB (A) or qPCR (B). (C and D) DC2.4 cells were treated with clotrimazole or DMSO, then transfected with OVA (100 µg/mL) for 20 hours, then co-culture with B3Z for an additional 24 hours, after which B3Z cell activation was measured by LacZ activity and IL-2 production (C), the expression of Chop was detected by WB and qPCR (D). (E) DC2.4 cells were treated with clotrimazole or DMSO for 24 hours, after which the expression of CD86 was detected by flow cytometry and qPCR. (F) DC2.4 cells were treated with DMSO, clotrimazole, bafilomycin or clotriamzole +bafilomycin for 24 hours, after which the protein level of CHOP was detected by WB. (G) DC2.4 cells were treated with DMSO or clotrimazole for 24 hours, after which the protein level of CHOP was detected by WB. (H) DC2.4 cells were treated with DMSO, clotrimazole, lactate or clotriamzole +lactate for 24 hours, after which the protein level of CHOP was detected by WB. Data in A, C, E–H are the representative result of three repeated experiments. Data in (B, D and E) are shown as mean±SD of three replicates from one representative experiment. *p<0.05, **p<0.01, ***p<0.001, by one-way analysis with Bonferroni’s post-test. OVA, antigen ovalbumin; CLT, clotrimazole; DMSO, dimethyl sulfoxide; WB, Western blot; SCR, scramble; CHOP, C/EBP homologous protein; NT, no lactate treatment.