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Vgll4b and Vgll4l are sufficient to rescue the loss of Vgll4 activity in the pLLP, and Vgll4b TDU2 is required for this function. MIP of confocal Z-stacks showing the pLLP in vgll4b+/+;vgll4l+/+ (a), vgll4b−/−;vgll4l+/+ (c), vgll4b−/−;vgll4l−/− (e) embryos and the same genotypes injected with tRFP-vgll4b mRNA (b, d, f). g Quantification of pLLP cell number in each condition (N ≥ 2 independent replicates). MIP of confocal Z-stacks showing the pLLP in vgll4b+/+;vgll4l+/+ (h), and vgll4b−/−;vgll4l−/− (i) embryos, injected with vgll4l mRNA at 100 ng/ μL (j, k) or at 200 ng/μL (l, m). n Quantification of pLLP cell number in each condition (N ≥ 2 independent replicates). MIP of confocal Z-stacks showing the pLLP in vgll4b+/+;vgll4l+/+ (o), and vgll4b−/−;vgll4l−/− (p) embryos, injected with tRFP-vgll4bΔTDU1 (q, r) or tRFP-vgll4bΔTDU2 (s, t) mRNA. u Quantification of pLLP cell number in each condition (N ≥ 3 independent replicates). Data are presented as mean ± SD. Unpaired t-tests (Mann–Whitney) were conducted. Scale bar: 20 μm (a–f, h–m, o–t).
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