Journal: Frontiers in Cell and Developmental Biology

Article Title: Ppp2r2a Knockout Mice Reveal That Protein Phosphatase 2A Regulatory Subunit, PP2A-B55α, Is an Essential Regulator of Neuronal and Epidermal Embryonic Development

doi: 10.3389/fcell.2020.00358

Figure Lengend Snippet: Defective epidermal barrier in Ppp2r2a knockout embryos is associated with changes in proliferation, and p-cJun signaling. (A) X-gal substrate penetration (permeability) assay in E18.5 embryos. Areas stained blue on the Ppp2r2a –/– embryo indicates regions with an impaired epidermal barrier. White arrowheads mark regions (umbilical cord and tail) that stained blue as they were cut during dissection. Representative of n = 3. Scale bar = 1 cm. (B) Ppp2r2a + / + and Ppp2r2a –/– epidermal regions at E14.5 and E18.5 showing Ki67 (proliferation marker) staining. Dashed black line indicates the dermal-epidermal junction. (C) Quantitation of Ki67 positive and negative cells in E14.5 and E18.5 epidermis. Ki67 + cells were significantly increased in the Ppp2r2a –/– suprabasal epidermis compared to the wild-type. * p < 0.05, unpaired two-tailed t -test. (D) IHC for p-cJun (S63) in Ppp2r2a + / + and Ppp2r2a –/– epidermal regions. Dashed black line indicates the dermal-epidermal junction. All scale bars = 50 μm.

Article Snippet: Primary antibodies were rabbit polyclonal anti-cleaved caspase 3 (1:200; Cell Signaling Technology #9661), rabbit monoclonal anti-Ki67 [SP6] (1:100; Abcam, ab16667) and rabbit monoclonal p-cJun (1:100; Cell Signaling Technology #2361).

Techniques: Knock-Out, Permeability, Staining, Dissection, Marker, Quantitation Assay, Two Tailed Test