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. Author manuscript; available in PMC: 2014 Dec 1.
Published in final edited form as: J Invest Dermatol. 2013 Dec 10;134(6):1710–1717. doi: 10.1038/jid.2013.530

Figure 4. Neutralization of IL-12 and IL-23 inhibits the repair of BMDC in TLR4 deficient mice.

Figure 4

BMDC were prepared as described in the Methods section. BMDC from TLR4 deficient mice were exposed to acute UVB (3 mJ/cm2) radiation and thereafter harvested at 0-24 h. (a) Neutralizing anti-IL-12p35 antibody was added to BMDC cultures before UV exposure. BMDC were exposed to acute UVB (3 mJ/cm2) radiation and thereafter harvested at 0-24 h and analyzed for CPD. BMDC were subjected to immunofluorescent staining to detect CPD+ cells that are green as mentioned in the Methods section. CPD were not detected in non-UV-exposed BMDC. (b) CPD+ cells were counted as a percentage of BMDCs from TLR4-/- mice. There were significantly more CPD at 24h post UV-exposure in the BMDC that were treated with neutralizing anti-IL-12p35 antibody than those treated with control IgG. (c) There was a decrease in XPA mRNA expression after neutralization with anti-IL-12p35 antibody as determined by quantitative real time PCR (qPCR) in the BMDC of TLR4-/- mice. (d) Neutralizing anti-IL-23p19 antibody was added separately to BMDC cultures before UV exposure. BMDC were exposed to acute UVB (3 mJ/cm2) radiation and thereafter harvested at 0-24 h and analyzed for CPD. BMDC were subjected to immunofluorescent staining to detect CPD+ cells that are green as mentioned in the Methods section. CPD were not detected in non-UV-exposed BMDC. (e) CPD+ cells were counted as a percentage of BMDCs from TLR4-/- mice. There were significantly more CPD at 24h post UV-exposure in the BMDC that were treated with neutralizing anti-IL-23p19 antibody than those treated with control IgG. (f) There was a decrease in XPA mRNA expression after neutralization with anti-IL-23p19 antibody as determined by quantitative real time PCR (qPCR) in the BMDC of TLR4-/- mice. Experiments were conducted and repeated separately in triplicates in each group with identical results. Scale bar = 50 μm. The experiments were repeated twice with similar results. (*p<0.05, **p<0.01 and ***p<0.001).