Figure 1. STING is correlated with DNA damage and affects neural progenitor cells. A) Images of cerebral cortex sections of E14, E17, and P0 labeled for 𝛾H2AX and DAPI. The bar graph shows the number of 𝛾H2AX+ cells in the VZ/SVZ per 8 × 105 μm2. SVZ, subventricular zone; VZ, ventricular zone; IZ, intermediate zone; CP, cortical plate. Scale bar, 100 μm. B) Western blot analysis of 𝛾H2AX, STING, AIM2, and TLR9 in neural progenitor cells (NPCs) after treatment with bFGF of various concentrations for 16 h in proliferation medium (suspension culture) (n = 4 independent experiments).
C,D) NPCs were isolated at E12.5 and then treated with bFGF (0.1, 50 ng mL−1) for 16 h in proliferation medium (suspension culture). Then cells were fixed and costained with anti-𝛾H2AX and anti-STING antibodies. Scale bar, 15 μm (C). D) Right, bar graph shows the fluorescence intensity of STING and 𝛾H2AX (n = 6 field of view of three individual experiments). E) STING is expressed in the development of the cerebral cortex. Cerebral cortex of different developmental stages (E10, E12, E14, E17, and P0) were isolated and lysed for Western blot analysis of STING, Tuj1, PAX6 (n = 3 individual experiments). F) STING knockdown results in abnormal cell position in embryonic cerebral cortex. Left, STING knockdown or control plasmids were electroporated into brain of E13.5 and analyzed at E16.5. Right, bar graph shows the percentage of GFP-positive cells in each region (n = 5 embryos from three different mothers). G) NPCs from E12.5 cerebral cortex were treated with 0.2 × 10−6 m cGAMP for 1 d before qRT-PCR analysis of neural differentiation-related gene mRNA and H) innate immune-related gene mRNA levels. I) NPCs were isolated from E12.5 cerebral cortex and cultured for 2 d. Then cells were treated with 0.2 × 10−6 m cGAMP for 1 d before fixed and costained with anti-SOX2 and anti-MAP2 antibodies. Bar graph shows the ratio of MAP2 or SOX2-positive cells in control and cGAMP treated groups (n = 4 independent experiments). Scale bar, 30 μm. Error bars represent means ± SEM; two-tailed unpaired t-test, n.s., not significant, *P < 0.05, **P < 0.01, or ***P < 0.001.
背景：
以前的研究已經報導了一些幹細胞，如胚胎幹細胞和造血幹細胞會發生DNA損傷，然而，對神經前體細胞的DNA損傷還不清楚。
𝛾H2AX是一種敏感的DNA損傷標誌物，主要在腦室區/室下區表達，在胚胎14d時高表達，並隨著發育的進展逐漸降低
鹼性成纖維細胞生長因子(BFGF)是一種廣為人知的鼻咽癌增殖神經營養因子
目的和方法：
為了進一步確定DNA損傷是否主要存在於快速增殖的NPC中，我們在含有不同濃度bFGF的增殖培養基中培養了原代NPC
結果：
組蛋白𝛾H2AX的磷酸化水平與bFGF濃度呈正相關
經兩種DNA損傷劑(DDA)處理後，在NPC中檢測到更多的胞漿DNA
結論：
篩選出三個主要的DNA活化分子：STING、AIM2和TLR9。我們發現，STING在鼻咽癌中的表達與𝛾H2AX呈正相關，提示STINT可能在鼻咽癌中起重要作用。
目的和方法：
為了探討STING在NPC中的作用，我們首先檢測了STING在胚胎10~P0的胚胎皮質中的表達
為了進一步研究刺痛在腦發育過程中是否在神經發生中發揮獨特的作用，構建敲低細胞系
方法：
利用宮內電穿孔技術(IUE)將SING-shRNA導入妊娠小鼠E13胚胎皮質中的神經前體細胞
CGAS是一種研究較多的刺激物上遊分子