Extended Data Figure 1: Myt1l antibody design and characterization.

a, Schematic of mouse Myt1 family members mmMyt1 (Q8CFC2), mmMyt1l (P97500) and mmSt18 (A5LFV3) as well as human hsMyt1l homologue (Q9UL68). Highlighted are the nuclear localization signals (NLS), aspartic acid/glutamic acid-rich (Asp/Glu-rich), serine-rich (Ser-rich), Myt1 and coiled-coil domains and the CCHC-type zinc-fingers (ZF). Also shown is the predicted antigenicity and the conservation between the proteins generated using EpiC and T-Coffee, respectively. Based on these data, a fragment of mmMyt1l (amino acids 171–420) was used as an antigen for antibody induction in rabbits. The sequence identities among the antigen regions and the full-length proteins and their molecular masses are shown (right). b–d, Western blots of MEF cells upon induction of Flag-tagged mmMyt1l (b), HEK293 cells upon transfection of Flag-tagged mmMyt1, mmMyt1l, St18, and untagged hsMyt1l (c), and E13.5 embryonic mouse whole brain lysate using preimmune serum and antibodies against Myt1l, Flag, and tubulin (d). e, Microscopy images of HEK293 cells upon transfection of Flag-tagged mmMyt1l followed by immunofluorescence using antibodies against Flag (red) and Myt1l (green). f, Microscopy images of a section from adult mouse cortex upon immunofluorescence using antibodies against NeuN (red) and Myt1l (green), and DAPI staining (blue). Scale bar, 10 μm. g, Myt1l antibody specifically marks mouse brain neurons in vivo. Immunofluorescence microscopy images of adult mouse brain cortex sections using antibodies against neuron-specific NeuN and Map2 or oligodendrocyte-specific Olig2 and Apc. Astrocytes (Gfap) and microglia (Iba1) are shown in red with Myt1l (green) and DAPI staining (blue). Note that Myt1l overlaps only with neuronal markers. Scale bar, 20 μm.