Research Paper Volume 13, Issue 6 pp 9011—9027

Functional genomics study of protein inhibitor of activated STAT1 in mouse hippocampal neuronal cells revealed by RNA sequencing

Overexpression of Pias1 regulates AD marker gene expression and cell proliferation in HT-22 cells. (A) qRT-PCR analysis of Pias1 mRNA expression in the hippocampal neuronal HT22 cells transfected with the indicated transgenes. Data represent the mean ± SD of three biological replicates. *p **p B) Western blot analysis of PIAS1 in the hippocampal neuronal HT22 cells with stable Pias1 transgenic expression. (C) Cell proliferation was determined by CCK-8 assays in HT-22 cells transfected with PiggyBac vector (PB) or PB-Pias1. Data represent the mean ± SD. *p **p D) qRT-PCR analysis of Neurod1, NeuN, Mapk2, Gsap, Mapt, App, Bace1 and Bace2 expression levels in PB and PB-Pias1 hippocampal neuronal HT22 cells. Data are presented as the mean ± SD of three independent experiments. *p **p E) Immunofluorescence staining of APP in the hippocampal neuronal HT22 cells overexpressing PB-Pias1. Bar: 100 μm.

Figure 2. Overexpression of Pias1 regulates AD marker gene expression and cell proliferation in HT-22 cells. (A) qRT-PCR analysis of Pias1 mRNA expression in the hippocampal neuronal HT22 cells transfected with the indicated transgenes. Data represent the mean ± SD of three biological replicates. *p < 0.05, **p < 0.01 versus PB vector. The control check (CK) group represents the baseline expression of Pias1 in HT-22 cells. (B) Western blot analysis of PIAS1 in the hippocampal neuronal HT22 cells with stable Pias1 transgenic expression. (C) Cell proliferation was determined by CCK-8 assays in HT-22 cells transfected with PiggyBac vector (PB) or PB-Pias1. Data represent the mean ± SD. *p < 0.05, **p < 0.01. (D) qRT-PCR analysis of Neurod1, NeuN, Mapk2, Gsap, Mapt, App, Bace1 and Bace2 expression levels in PB and PB-Pias1 hippocampal neuronal HT22 cells. Data are presented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01. (E) Immunofluorescence staining of APP in the hippocampal neuronal HT22 cells overexpressing PB-Pias1. Bar: 100 μm.