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Fig. 3 | Fluids and Barriers of the CNS

Fig. 3

From: Streptococcus agalactiae disrupts P-glycoprotein function in brain endothelial cells

Fig. 3

Effects of GBS infection on P-gp expression in BECs. a BECs were infected with GBS (MOI = 10) for 5 h, and protein lysates were collected. Western blot analysis was performed on lysates using COXIV as a loading control, representative blot shown. b Densitometry was conducted to determine fold change in P-gp expression using FIJI ImageJ software after normalization to respective COXIV bands on all experiments performed in triplicate on three independent differentiations (n = 9). c Representative flow cytometry histogram of BECs stained for total P-gp after 5 h of GBS infection (MOI = 10) compared with uninfected BECs (Control). d P-gp abundance is plotted as the median fluorescence intensity generated by flow cytometry normalized to uninfected control on all experiments performed at least in duplicate on three independent differentiations (n = 7). e qPCR performed for ABCB1 (P-gp) and normalized to 18S rRNA for BECs with or without GBS infection at an MOI of 10 for 5 h. Data presented are from three experiments performed in triplicate. f Quantitation of relative P-gp pixel intensity overlayed with vascular lectin staining in mouse brains. Control mice (n = 5) with 12 FOV had 481 lectin positive vessels analyzed, GBS mice (n = 6) with 12 FOV had 544 lectin positive vessels analyzed. g Representative images of mouse brains stained for lectin (green), P-gp (red), and DAPI (blue) after GBS infection. Control mouse with no GBS in the brain (top), and mouse with GBS in the brain (bottom). Scale bar represents 50 μm. Data represent mean ± SD. ***p < 0.001; Student’s t test

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