Figure 2.

Localization of Lunapark in Mice and Humans

(A) Graph showing the expression of mouse Lnpk in relation to the geometric mean of three housekeeping genes via quantitative RT-PCR. Time points range from embryonic day 12.5 (E12.5) to postnatal day 0 (P0). Graph shows mean ± SEM for each time point. n = 3 animals.

(B) Image of in situ hybridization using RNAscope against the Lnpk mRNA transcript. Shown are DAPI and the in situ signal. B′ shows the Lnpk signal alone for better visualization. Abbreviations are as follows: VZ, ventricular zone; SVZ, subventricular zone; IZ, intermediate zone; CP, cortical plate; and MZ, marginal zone.

(C) Image shows an H9-derived NPC stained with DAPI and antibodies against LNP and SOX2. C′ and C″ show gray-scale images of the indicated channels.

(D–G) Images show an H9-derived NPC, driven toward differentiation by 1 day of bFGF withdrawal, stained with DAPI and antibodies against LNP and TUBB3 (Tuj). Shown are an overview (D), magnification of the soma (E), magnification of a process branch point (F), and magnification of the tip of the extending process (G). In addition to the merged image (E–G), gray-scale images of the LNP and TUBB3 channels are shown (E′–G′ and E″–G″, respectively). LNP appeared to be upregulated in differentiating NPCs and also accumulated in branch points and the tip of the extending neurite that was reminiscent of an axonal growth cone. The inlay in (F′) shows the boxed area with higher contrast for better visualization of the increased signal of LNP at the branching point. Scale bars represent 50 μm (B), 10 μm (C and G″), and 25 μm (D).