Figure 1 | Scientific Reports

Figure 1

From: The distinct effects of P18 overexpression on different stages of hematopoiesis involve TGF-β and NF-κB signaling

Figure 1

Involvement of P18 is involved in inhibitory effects on the mesoderm–hemogenesis transition by RUNX1b, and establishment of inducible P18 transgenic hESC lines. D0-induced RUNX1b/hESCs (a) were co-cultured with AGM-S3 cells at D4. (b) qRT-PCR analysis and (c) Western blotting (WB) analysis revealed that when DOX was added from D0, P18 was upregulated at the mRNA and protein levels at D4, and that these effects can be counteracted by addition of 0.33 μM RepSox from D0. (d) Schematic diagram of the piggyBac vector used to induce P18 overexpression. TRE, tet-on regulation element; CMV Mini, minimum promoter of cytomegalovirus; T2A, Thosea asigna virus 2A peptide. (e) After P18/hESCs was treated with DOX for 48 h, co-expression of GFP was observed by fluorescence microscopy. (f) qRT-PCR analysis and (g) WB analysis confirmed that inducible expression of P18 was highly stringent and effective at the mRNA and protein levels. GAPDH was used as an internal control. (h) WB analysis with anti-SOX2, -OCT4, and -NANOG antibodies confirmed the normal pluripotency of P18/hESCs. The loading control was GAPDH. All results are expressed as means ± SD of three repeated experiments, and p < 0.05 was considered significant (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

Back to article page