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. 2021 Aug 5;17(8):e1009686. doi: 10.1371/journal.pgen.1009686

Fig 2. TUBL expression promotes cell cycle progression and inhibits differentiation in human keratinocytes.

Fig 2

(A, B) GSEA plots for gene sets related to the cell cycle (A) or keratinocyte differentiation (B) constructed from RNA-seq data obtained for NHEKs expressing GFP or human TUBL. FDR, false discovery rate; NES, normalized enrichment score. (C) Summary of GSEA results for gene sets related to the cell cycle or keratinocyte differentiation. NOM, nominal. (D) Flow cytometric traces and quantification of BrdU incorporation for NHEKs stably expressing GFP or TUBL. Data in the right panel are means ± SD (n = 3 independent experiments). ***p < 0.005 (Student’s t test). (E) RNA-seq results for differentially expressed genes related to basal, early, or late keratinocyte differentiation. The results for NHEKs expressing TUBL were normalized by those for NHEKs expressing GFP. *p < 0.05, ***p < 0.005; NS, not significant (adjusted p values). (F, G) RT-qPCR analysis of TINCR and involucrin (IVL) mRNA abundance in NHEKs transfected with the indicated control or TINCR siRNAs for 1 day and then subjected to calcium-induced differentiation for 0 or 6 days (F). The results for TINCR abundance at day 0 of differentiation are expanded in (G). Data are means ± SD (n = 3 independent experiments). ***p < 0.005, NS (Student’s t test). (H) Quantification of BrdU incorporation for NHEKs transfected with the indicated control or TINCR siRNAs for 2 days. Data are means ± SD (n = 3 independent experiments). **p < 0.01, ***p < 0.005 (Student’s t test).