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. 2016 Nov 14;8(12):1455–1469. doi: 10.15252/emmm.201606623

Figure 2. Scheme of SHOX and CYP26C1 proteins and luciferase assays.

Figure 2

  1. SHOX protein representation and luciferase assay. The variants found in SHOX are indicated in red. Luciferase assays to test the impact of SHOX mutations on its transcriptional activity were performed on the FGFR3 promoter in U2OS cells (n = 4). Experiments were performed in triplicates. pcDNA4‐TO empty vector was used as control. R153L represents a mutation known to affect SHOX protein activity and was used as a positive control (Schneider et al, 2005). Homeodomain, DNA binding domain; SH3, Src homology 3 domain; OAR, OtpAristalessRax domain; RLU, relative light units.
  2. CYP26C1 protein representation and luciferase assay. Variants found in CYP26C1 are indicated in green. Cignal RARE system luciferase assays to test the impact of CYP26C1 variants on its RA degradation activity were performed in U2OS cells treated with 250 nM all‐trans retinoic acid (ATRA) for 24 h (n = 4). Experiments were performed in triplicates. pIRES2‐EGFP empty vector was used as control. The residue C459 represents the iron binding residue (Q6VOL0, UniProtKB) and was mutated to Ala and used as a positive control. TM, transmembrane helix; P450, cytochrome p450 domain. RLU, relative light units.
Data information: The box represents the interquartile range. The whiskers represent Min to Max. *P‐value = 0.0286, two‐tailed Mann–Whitney non‐parametric t‐test.