Figure 2.

Robust and chronic visualization of blood plasma by in vivo transgene expression of Alb-mNG in hepatocytes

(A) Schematic of approach for the in vivo experiments. AAV8-P3-Alb-mNG is administrated to mice via i.p. or i.v. injection (left). Alb-mNG expression was monitored by collecting a blood sample from the tail. Brain and liver tissues and blood were collected for morphological and biochemical examination (right).

(B) Example of the fluorescence signals in blood samples collected on days 2 and 5 from a mouse that received i.p injection of AAV8-P3-Alb-mNG.

(C) Plasma Alb-mNG fluorescence intensity over a time course of 8 weeks. Significant effect of time (one-way ANOVA: p < 0.05) (n = 6 mice).

(D) Mouse liver images after 3 weeks of AAV8-P3-eGFP (positive control), PBS (negative control), and AAV8-P3-Alb-mNG i.p. injection. Scale bars, 500 μm.

(E) mNG concentration in plasma samples (n = 3 mice). No significant effect of time (one-way ANOVA: p > 0.05)

(F) Plasma albumin concentration using albumin ELISA in PBS- (gray, n = 2) and Alb-mNG-injected (green, n = 3) mice.

(G) Plasma CRP levels for control or Alb-mNG i.p. injected mice during the 8 weeks of post-injection period (n = 6–12 mice).

(H) Example images of liver (top panel) and brain slices (bottom panel) of control- (PBS) or Alb-mNG-expressing mice immunostained for macrophages (liver) or microglia (brain) by IBA1 (purple) and DAPI (yellow). Brain sections of lipopolysaccharide (LPS)-injected mice displayed reactive microglia morphology, while resting microglia are observed in the Alb-mNG mouse. Scale bars, 10 μm.

Graphs show means ± SEM and individual values (except from C); ∗p < 0.05.