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. 2021 Nov 16;9:779373. doi: 10.3389/fcell.2021.779373

FIGURE 8.

FIGURE 8

NFATc3 is a key transcription factor in the regulation of cell cycle by Ttyh1. (A) RT-qPCR analysis showed the expression levels of five NFAT family members between control and Ttyh1 KO neurospheres at primary culture (P0) and Passage 2 (P2). The RNA expression of NFATc3 increased at P0 and decreased significantly at P2. (B) Western blotting analysis showed the protein level of NFATc3 was also increased at P0 but decreased at P2. (C) Bright field microscopy showed the morphological disparity among NSCs transfected with negative control shRNA (NC), shTtyh1, and shTtyh1+ NFATc3 (rescue group), respectively. After 48 or 72 h of transfection, the results showed that overexpression of NFATc3 partially rescued the phenotypic changes caused by knockdown of Ttyh1. (D) RT-qPCR analysis showed that when Ttyh1 was knockdown, Ccnd1 was significantly increased, and p21 was slightly increased too. When NFATc3 was overexpressed at the same time, p21 increased significantly, whereas Ccnd1 showed a down-regulated trend. Scale bar = 100 μm in C. n ≥ 4 for all experiments. Data are expressed as mean ± SEM. Statistical significance was calculated using an unpaired, two-tailed Student’s t-test. (∗∗∗∗) p < 0.0001, (∗∗∗) p < 0.001, (∗∗) p < 0.01, (∗) p < 0.05.