(A–C) E13 murine SMG+SLG cultured for 48 hr ± parasympathetic ganglion (nerves) and subjected to immunofluorescent analysis (A). Glands were immunostained for markers of duct cells (KRT19, KRT7), SOX2, SOX10+ acinar progenitors, AQP5+ acinar cells and epithelial cells (E-cadherin; ECAD). The number of acini (B) and AQP5+ and SOX10+ cells (C) were quantified. (D–F) E11.5 murine SMG+SLG deficient in Phox2b were cultured for 60 hr. Glands were were immunostained for markers of nerves (TUBB3), acinar progenitors (SOX10), basal epithelial progenitors (KRT5) and epithelial cells (E-cadherin; ECAD; D), the number of acini were quantified (E) or qPCR was performed. (F; n = 3 embryos per genotype). Scale bars = 200 µm in (A), left panels and (D), left panels; 50 µm in (A), right panels and (D), middle and right panels; 20 µm in (A), middle panels. Data in B, C and E are means ± s.d of three biological replicates and three experiments and were subjected a Student’s t-test. *p<0.05 **p<0.01. Data in F are means+s.d. and were analyzed using a one-way analysis of variance with post-hoc Dunnett’s test. *p<0.05, **p<0.01.