FIGURE SUMMARY
Title

Cloning of nine glucocorticoid receptor isoforms from the slender African lungfish (Protopterus dolloi)

Authors
Katsu, Y., Oana, S., Lin, X., Hyodo, S., Bianchetti, L., Baker, M.E.
Source
Full text @ PLoS One
Fig. 1

Total RNA was isolated from P. dolloi ovary and translated into cDNA. PCR was performed using four primer sets based on the sequence of P. annectens GR, as described in the Methods section. The amplified DNA fragments were sub-cloned into a vector for sequence analysis. Similar to other steroid receptors, slender African lungfish GR can be divided into four functional domains [6, 8], consisting of a ligand-binding domain (LBD) at the C-terminus, a DNA-binding domain (DBD) in the center that is joined to the LBD by a short hinge domain (hinge), and a domain at the amino-terminus (NTD). GenBank accession no. BDF84376 for GR-A1, BDF84377 for GR-A2, BDF84378 for GR-B1, BDF84379 for GR-B2, BDF84380 for GR-B3, BDF84381 for GR-C1, BDF84382 for GR-C2, BDF84383 for GR-C3, and BDF84384 for GR-C4. Sequences were aligned with Clustal W [35], as described in the Methods section.
Fig. 2

Fig 2. Multiple alignment of the amino acid sequences of three African lungfish GRs and four West African lungfish GRs.

West African lungfish glucocorticoid receptor sequences were downloaded from GenBank (Accessions XP_043925084 for X1, XP_043925085 for X2, XP_043925087 for X3, XP_043925088 for X4). Sequences were aligned with Clustal W [35], as described in the Methods section.
Fig. 3

Fig 3. Multiple sequence alignment of slender African lungfish GR, West African lungfish GR, coelacanth GR, zebrafish GR and human GR.

Glucocorticoid receptor sequences were downloaded from GenBank (Accession no. NP_000167 for human GR, XP_005996162 for coelacanth GR, and NP_001018547 for zebrafish GR) and aligned with Clustal W [35], as described in the Methods section. The NTD in zebrafish GR has gaps and sequence differences with the other GRs.
Fig. 4

Fig 4. Comparison of functional domains of slender lungfish GR with domains in West African lungfish GR, coelacanth GR, zebrafish GR, human GR.

Comparison of domains in slender African lungfish GR with GRs from West African lungfish, coelacanths, humans and zebrafish and MRs from slender African lungfish, West African lungfish, humans and zebrafish. The functional NTD (A/B), DBD (C), hinge (D) and LBD (E) domains are schematically represented with the numbers of amino acid residues and the percentage of amino acid identity depicted.
Fig. 5

Fig 5. Phylogeny of slender African lungfish glucocorticoid receptors, West African lungfish glucocorticoid receptors, coelacanth GR and elephant shark GR. MEGA5 [36] was used to construct this phylogeny. Statistics are based on 1,000 runs.

Fig. 6

Fig 6. Location of PCR primers used for cloning of slender African lungfish GR, coelacanth GR, elephant shark GR, zebrafish GR and human GR.

The correct 1st/2nd-reverse primer for PCR cloning of P. dolloi GR is WQRFFQ instead of WQRFYQ.
Acknowledgments
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