Extended Data Figure 4: KBTBD8 is required for differentiation into functional neural crest cells.
From: Cell-fate determination by ubiquitin-dependent regulation of translation
a, H1 hESCs stably expressing control or KBTBD8 shRNAs were subjected to neural conversion for 43 days and analysed by immunofluorescence microscopy against GFAP (glia), smooth muscle actin (SMA; mesenchymal cells), and neurofilament L (neurons). b, Control H1 hESCs or hESCs depleted of KBTBD8 were subjected to neural conversion for 43 days and expression of markers for glia (GFAP), mesenchyme (smooth muscle actin, SMA), melanocytes (TYRP1, DCT), chondrocytes (COL2A1), or CNS derivatives (PAX6, NESTIN, neurofilament L) was analysed by qRT–PCR (mean of 3 technical replicates ± s.e.m.). c, Xenopus tropicalis embryos were injected at the two-cell stage with splice-blocking morpholinos (sMO) against CUL3 or KBTBD8, or with a dominant-negative construct of CUL3 that allows KBTBD8 to bind, but not ubiquitylate, substrates. Neural crest formation was monitored by SOX10 in situ hybridization. Quantification included experiment shown in Fig. 1d (mean of 3 biological replicates ± s.d.; ∼20 embryos per condition and replicate).
