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Fig. 7 | Stem Cell Research & Therapy

Fig. 7

From: Long noncoding RNA CCDC144NL-AS1 knockdown induces naïve-like state conversion of human pluripotent stem cells

Fig. 7

Rescued knockdown of CCDC144NL-AS1 in CCDC144NL-AS1-KD hESCs and the signaling pathway state of CCDC144NL-AS1-KD hPSCs. a Morphologies of CCDC144NL-AS1-KD-H9 clusters before and after expression of CCDC144NL-AS1 was rescued. Scale bars, 100 μm. b qPCR validation of CCDC144NL-AS1’s rescued expression in CCDC144NL-AS1-KD-H9 cells. CCDC144NL-AS1-KD-H9 cells infected with empty overexpression viruses were used as control cells. Error bars indicate SEM (n = 3). ***p < .001. c Western blot analyses of naïve pluripotency-related NANOG protein levels in CCDC144NL-AS1-rescued CCDC144NL-AS1-KD-H9 cells and CCDC144NL-AS1-KD-H9 cells infected with empty overexpression viruses were used as control cells. d Western blot analyses of total and phosphorylated protein levels of MAPK in CCDC144NL-AS1-KD-H9 and NC-H9 cells. e Heatmap of genes involved in MAPK/ERK signaling pathway in CCDC144NL-AS1-downregulated hPSCs and control cells. f Western blot analyses of DUSP3, DUSP6, DUSP5, and DUSP7 protein levels in CCDC144NL-AS1-KD-H9 and NC-H9 cells. g Western blot analyses of total and unphosphorylated (active) β-catenin protein levels in CCDC144NL-AS1-KD-H9 and NC-H9 cells. h Heatmap of genes involved in the Wnt signaling pathway in CCDC144NL-AS1-downregulated hPSCs and control cells. i Western blot analyses of total SMAD2/3 and phosphorylated SMAD2 protein levels in CCDC144NL-AS1-KD-H9 and NC-H9 cells. j Heatmap of genes involved in the TGF-β signaling pathway in CCDC144NL-AS1-downregulated hPSCs and control cells. The expression levels of genes in heatmaps are determined by RNA-seq data. β-Tubulin was used as an endogenous control in all western blot analyses

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