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. 2020 Jul 21;11(4):e01249-20. doi: 10.1128/mBio.01249-20

FIG 7.

FIG 7

Three lead compounds prevent C. neoformans (Cn) from crossing the BBB. (A) Transcytosis assays 6 h posttreatment and postinoculation in a 96-well format of the in vitro model of the BBB (lower, right diagram). Lead compounds were tested in transcytosis assays with four fungal strains: wild-type Sc (ScWT), Sc<CnMPR1> (expression of CnMPR1 cDNA in Sc), the Cnmpr1Δ deletion strain, and the isogenic wild-type H99 Cn strain. A dose of 5 μM of each lead compound was added to the upper well. Treatment with lead compounds (NP001-1C11, NP002-1D9, and NP003-2B11) prevents crossing of Cn compared to DMSO control (n = 7, per compound). *, P < 0.05, **, P < 0.01, ***, P < 0.001; ns, not significant. (B) Compounds did not inhibit growth of Cn (P > 0.05). (C) Integrity of BBB was monitored by FITC-dextran (70-kDa) permeability assays in the in vitro BBB model. In the absence of hCMEC/D3 cells, FITC-dextran was able to freely cross the transwell (blue bar) but unable to cross when hCMEC/D3 cells were present and the BBB was intact (red bar). Treatment with compounds and addition of different strains did not promote FITC crossing, suggesting that the integrity of the BBB was not damaged by the three lead compounds.