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Fig. 1
The highly conserved gene letm1 is broadly expressed in zebrafish with high levels in brain. (A) Phylogenetic analysis of Letm1/2 family proteins using the free online access tools phylogeny.fr and itol.embl.de/. HCCR1 was used as an out-group. (B, C, D) Confocal images of HeLa cells expressing CMV-Drletm1-eGFP (green) and stained with MitoTracker Red (magenta). Scale bar 10 μm. (E) Relative mRNA levels of letm1 in zebrafish larvae, head, and tail at 3 and 7 day post fertilization (dpf), normalized to the eukaryotic elongation factor-1 α (eef1a). Each data point represents a biological replicate. 3 dpf head (n = 12); 3 dpf tail (n = 9); 7 dpf head (n = 14); 7 dpf tail (n = 9). t test: ****P ≤ 0.0001. (F) Relative mRNA levels of letm1 in adult organs (SB, swimming bladder), normalized to eef1a. Each data point represents a biological replicate. Mean ± SEM. (G, H) In situ hybridization of letm1 on wild-type embryos at 1 dpf. Scale bar 1 mm. (I) Schematic representation of the adult zebrafish brain in lateral view. Blue vertical lines represent the location of the brain in lateral view. Blue vertical lines represent the location of the brain slices displayed. OB, olfactory bulb; Tel, telencephalon; TeO, optic tectum; Ce, corpus cerebelli; HT, hypothalamus; IL, inferior lobe; MO, medulla oblongata. (J, K, L) In situ hybridization of letm1 on coronal slices of wild-type adult brain. (J’, K’, L’) Magnification of the left panel. (M, N, O) Sense control. Scale bars 500 μm.