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. Author manuscript; available in PMC: 2021 Jan 13.
Published in final edited form as: Stem Cell Res. 2020 Oct 15;49:102048. doi: 10.1016/j.scr.2020.102048

Fig. 4. Transcriptional analysis of cells at each phase of the spheroid re-plating assay.

Fig. 4.

(A) Schematic illustrating the workflow employed to determine the gene expression profile of NHKc at different stages of the epidermal spheroid re-plating assay. (B) Phase contrast images of a (B1) 2-D monolayer mass cultured NHKc colony newly isolated from a skin explant, (B2) its corresponding 3-D suspension spheroid, and (B3) the subsequent spheroid-derived cultures obtained upon transfer of 3-D spheroids into 2-D monolayer culture. (B4-B6) corresponding cultures immunostained against Keratin 14. White contour lines indicate spheroid edge. (C) The expression of mRNAs encoding epidermal basal genes: K15, K5, K14; (D) Suprabasal differentiation genes: filaggrin, involucrin, K10; (E) epidermal proliferation genes: pan-P63, delta Np63, Ki-67; and (F) reprogramming genes: Nanog, Oct4, Sox2, Klf4 in suspension spheroids and SD cultures, relative to corresponding monolayer mass cultures as determined by RT-qPCR.