Figure 6
Figure 6. VLP, HIV-1, and exosome capture can be inhibited when particles are produced from ceramide-deficient cells. (A) A total of 105 mDCs were exposed to 2500 pg of p24Gag VLPHIV-Gag-eGFP produced from either FB1, NB-DNJ, or mock-treated HEK-293T cells, fixed, and analyzed by FACS to measure the percentage of eGFP-positive cells. mDCs captured significantly greater amounts of VLPHIV-Gag-eGFP produced from mock treated HEK-293T cells (P = .0003, paired t test). Mean values and SEM from 3 independent experiments, including cells from 5 donors are plotted. (B) mDCs were exposed to 10 ng of HIV-1Lai or HIV-1NL43 p24Gag produced from either FB1, NB-DNJ, or mock-treated HEK-293T cells, washed thoroughly to remove unbound particles, lysed, and assayed to measure the cell-associated p24Gag content by an ELISA. mDCs captured greater amounts of HIV-1 produced from mock-treated HEK-293T cells. Mean values and SEM from 2 independent experiments and cells from 3 donors are plotted. (C) mDCs (105) were exposed to 250 μg of ExosomesDiI produced from either FB1, NB-DNJ, or mock-treated Jurkat cells, fixed, and analyzed by FACS to measure the percentage of DiI-positive cells. mDCs significantly captured greater amounts of ExosomesDiI produced from mock-treated Jurkat cells (P = .0042, paired t test). Mean values and SEM from 3 independent experiments and cells from 6 donors are plotted.

VLP, HIV-1, and exosome capture can be inhibited when particles are produced from ceramide-deficient cells. (A) A total of 105 mDCs were exposed to 2500 pg of p24Gag VLPHIV-Gag-eGFP produced from either FB1, NB-DNJ, or mock-treated HEK-293T cells, fixed, and analyzed by FACS to measure the percentage of eGFP-positive cells. mDCs captured significantly greater amounts of VLPHIV-Gag-eGFP produced from mock treated HEK-293T cells (P = .0003, paired t test). Mean values and SEM from 3 independent experiments, including cells from 5 donors are plotted. (B) mDCs were exposed to 10 ng of HIV-1Lai or HIV-1NL43 p24Gag produced from either FB1, NB-DNJ, or mock-treated HEK-293T cells, washed thoroughly to remove unbound particles, lysed, and assayed to measure the cell-associated p24Gag content by an ELISA. mDCs captured greater amounts of HIV-1 produced from mock-treated HEK-293T cells. Mean values and SEM from 2 independent experiments and cells from 3 donors are plotted. (C) mDCs (105) were exposed to 250 μg of ExosomesDiI produced from either FB1, NB-DNJ, or mock-treated Jurkat cells, fixed, and analyzed by FACS to measure the percentage of DiI-positive cells. mDCs significantly captured greater amounts of ExosomesDiI produced from mock-treated Jurkat cells (P = .0042, paired t test). Mean values and SEM from 3 independent experiments and cells from 6 donors are plotted.

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