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. 2014 Jun 2;211(6):1063–1078. doi: 10.1084/jem.20132063

Figure 8.

Figure 8.

TNF and MMP9 participate in 12-HHT/BLT2–dependent mouse primary keratinocyte migration. (A) Relative mRNA levels of TNF were measured by Q-PCR in BLT2 WT and BLT2 KO mouse primary keratinocytes. Cells were stimulated with 100 nM 12-HHT for the indicated time (n = 4 experimental replicates). The relative mRNA level in unstimulated BLT2 WT keratinocytes was set as 1. (B) Relative levels of MMP9 mRNA were measured by Q-PCR in BLT2 WT and BLT2 KO mouse primary keratinocytes (n = 4 experimental replicates). The relative mRNA level in BLT2 WT keratinocytes was set as 1. (C) Mouse MMP9 protein levels in the culture medium of mouse primary keratinocytes were measured by ELISA after 12 and 24 h of culture (n = 4 experimental replicates). (D) The effect of mouse TNF neutralizing antibody D2H4 on the migration of BLT2 WT and BLT2 KO mouse primary keratinocytes. Cells were cultured in medium containing 1 nM 12-HHT and 10 ng/ml control IgG or 10 ng/ml D2H4 for 39 h (n = 5–6 experimental replicates). Data represent the mean ± SEM. **, P < 0.01; *, P < 0.05 (A and D, two-way ANOVA with Bonferroni post hoc tests; B and C, unpaired Student’s t test). All the results are representative of at least two independent experiments.