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. 2008 Dec 15;119(1):146–156. doi: 10.1172/JCI36851

Figure 3. Regulation of Raldh2 by Hoxd13.

Figure 3

(A) Luciferase reporter assay of a conserved region of murine Raldh2. Cos7 cells were transfected with Raldh2 reporter and Hoxd13 expression constructs carrying different Ala repeat length mutations (+7, +8, +9, +14). WT Hoxd13 activated Raldh2. Increasing length of the Ala repeat results in loss of activation. Error bars show SEM. EV, empty vector. (B) Quantitative PCR of chMM cultures showed upregulation of Raldh2 by WT Hoxd13. Infections with Hoxd13+7Ala and Hoxd13+14Ala were comparable with uninfected controls. Error bars show SD. (C) ChIP confirmed binding of WT Hoxd13 to the Raldh2 promoter. ChIP from chMM cultures was done with 4 different primer pairs. Real-time PCR with primer set 1 showed an approximately 45-fold enrichment of the Raldh2 promoter fragment precipitated with Hoxd13. Sets of 3 bars represent results from 3 independent experiments. Error bars show SD.