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Fig. 1 | Journal of Neuroinflammation

Fig. 1

From: SPARC expression by cerebral microvascular endothelial cells in vitro and its influence on blood-brain barrier properties

Fig. 1

SPARC expression is associated with proliferation of immature hCMEC/D3 and decreased as cells mature and reach confluence. a Confluency and monolayer intactness were visually assessed by phase contrast microscopy. b, c hCMEC/D3 cultures 20–30, 50–70, and 100 % confluent were analyzed for SPARC protein expression by immunoblotting. Data is pooled from two independent experiments, error bars represent standard error of the mean (SEM) with statistical significance by one-way ANOVA and Tukey’s comparison test **P < 0.001; ***P < 0.0001. d Representative confocal micrograph images show greater perinuclear and cytoplasmic SPARC staining (red) in subconfluent cultures compared to confluent cultures. Subconfluent cultures exhibit discontinuous peripheral ZO-1 bands (green), whereas confluent cells were tightly apposed and organized into monolayers with continual ZO-1 bands at interendothelial borders. Scale bar = 30 μm. e The individual cells in confluent monolayers express less SPARC than the cells in subconfluent cultures. Levels of SPARC immunoreactivity (negligible, low, moderate, or high) were assigned according to Table 1 (Additional file 1: Figure S1). Bars represent the average of results from n = 18 and n = 10 images enumerated for low, medium, or high levels in subconfluent (empty bar) vs. confluent (filled bar) cultures in replicate wells. f Summary of individual cell staining classification from both Fig. 1e (exp. A) and a second experiment (exp. B) with similar findings for the distribution of cells from (S) subconfluent vs. (C) confluent cultures

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