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. 2019 Jun 14;13:584. doi: 10.3389/fnins.2019.00584

FIGURE 3.

FIGURE 3

Preparation and identification of AQP4-RNAi-LV recombinants. (A) The schematic of the AQP4 siRNA sequence is shown. Enhanced red fluorescent protein (mcherryFP) as a reporter gene was inserted in the plasmid. The framework also contained the antibiotic ampicillin and pUC Ori promoters for vector expression. (B) qRT-PCR showed F1 was the most effective interference of AQP4. ∗P < 0.05 when compared to control group. (C) Fluorescent image of AQP4-RNAi-LV (red) transfected into 293T cells. (D) Immunohistochemistry shows the AQP4 immunoreactive products in the vector group and the AQP4-RNAi-LV group. Panel (D-a) shows AQP4 immunoreactive products in spinal cords of the vector group, and Panel (D-b) shows the AQP4 immunoreactive products in spinal cords of the AQP4-RNAi-LV group. It is clear that the degree of color in the vector group was stronger than the AQP4-RNAi-LV group, which suggests that the AQP4 interference affected the expression of AQP4 in vivo. Bar = 100 μm. (E) The area of neurons was measured by ImageJ, and the data showed that the area of neurons in the AQP4-RNAi-LV group was significantly decreased compared to the vector group at 7 days. ∗P < 0.05 when compared to vector group. (F) The BBB scores in the AQP4-RNAi-LV group were significantly higher than in the vector group at 5, 7, and 14 days (n = 6, P < 0.05). Values are expressed as mean ± SEM ∗P < 0.05 when compared to vector group.