Figure 4.

LUGGIGE in oral epithelia. (A) Expression of keratins in E13.5 pre-elevation palatal shelves (PS). K14 (in gray, Origene BP5009) is reduced in PS (indicated by arrowhead) as compared with nearby tongue and buccogingiva, whereas the oral keratin K6A (green, Biolegend Poly19057) and periderm marker K8 (red, Developmental Studies Hybridoma Bank TROMA-I) are present. (B, C) LUGGIGE. (B) Ultrasound image of lentiviral injection into the amniotic fluid surrounding E9.5 embryos. (C) LUGGIGE-transduced E17.5 embryo showing epithelial-specific expression of the nuclear histone H2B-mRFP1 reporter in epidermis and oral tissues. (D) Examples of lentiviral constructs harboring an shRNA and H2B-RFP1 reporter (left) for knocking down target genes, or Cre-RFP (right) for generating conditional knockouts. (E) Schematic of the palate region viewed by coronal section. (F–H) LUGGIGE can achieve high transduction in palatal cells at both early (F) and late (G, H) stages of palatogenesis. Scale bars: 100 µm. BG, buccogingiva; MES, medial epithelial seam; NS, Nasal septum; PS, palatal shelf; To, tongue.