Fig. 7
Expression of iNOS after LCN2 knockout and lentivirus transfection. a The WT and LCN2 knockout astrocytes were harvested 24 h after OGD for 0 h, 2 h, 4 h, 6 h, and 8 h. RT-PCR was used to detect the mRNA levels of LCN2. Compared with WT control, the LCN2 mRNA stayed no expression in LCN2 knockout astrocytes. The LCN2 mRNA level of WT astrocytes reaches up to over 10 times after 6 h OGD treatment. b RT-PCR was used to detect the mRNA levels of LCN2 after lentivirus overexpressed LCN2 transfection (LVLCN2) and its negative control (LVNC). The LVLCN2 treatment can increase the expression of LCN2 mRNA (P < 0.0001), and the LVNC had no effect on LCN2 expression (P > 0.05). c Astrocytes were harvested 24 h after 6 h OGD. Western blot was used to evaluate the protein iNOS level. The iNOS level increased sharply after OGD in WT astrocytes, but there was no iNOS expression in LCN2 knockout astrocytes with or without OGD treatment. d Astrocytes were transfected by LVLCN2 for 6 h and treated by OGD 24 h later. Astrocytes were harvested 24 h after 6 h OGD. The Western blot showed that anoxic induced iNOS expression in transfected astrocyte increased greatly. Antibody: rabbit anti-iNOS (1:1000, Cat#ab17945, Abcam), rabbit anti-beta-actin (1:2000, Cat#4970, Cell Signaling Technology), anti-rabbit (1:2000, Cat#14708, Cell Signaling Technology)