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Inhibition of EV secretion by PDDC and 5 in glial cells. Primary astrocytes were treated in parallel incubations with PDDC, 5, or cambinol in the 0.03–30 μM range as indicated; DMSO (0.02%) was used as vehicle control (set as 100% on y‐axis). Media was collected after 2‐hr incubation and centrifuged at 2,700 g for 15 min at 4°C. Supernatant was collected and the number of extracellular vesicles (EVs) was quantified using <t>ZetaView</t> Nanoparticle Tracker. The data points correspond to per cent inhibition of EV release (±SEM) over a concentration range for each compound; 100% release corresponds to 4.87 × 107 ± 0.098 EVs. Results are the average of three or four independent assays. Each independent assay was the average of three replicates. pEC50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] against response, using GraphPad Prism 7
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Particle Metrix zetaview nanoparticle tracker
Inhibition of EV secretion by PDDC and 5 in glial cells. Primary astrocytes were treated in parallel incubations with PDDC, 5, or cambinol in the 0.03–30 μM range as indicated; DMSO (0.02%) was used as vehicle control (set as 100% on y‐axis). Media was collected after 2‐hr incubation and centrifuged at 2,700 g for 15 min at 4°C. Supernatant was collected and the number of extracellular vesicles (EVs) was quantified using <t>ZetaView</t> <t>Nanoparticle</t> Tracker. The data points correspond to per cent inhibition of EV release (±SEM) over a concentration range for each compound; 100% release corresponds to 4.87 × 107 ± 0.098 EVs. Results are the average of three or four independent assays. Each independent assay was the average of three replicates. pEC50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] against response, using GraphPad Prism 7
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Inhibition of EV secretion by PDDC and 5 in glial cells. Primary astrocytes were treated in parallel incubations with PDDC, 5, or cambinol in the 0.03–30 μM range as indicated; DMSO (0.02%) was used as vehicle control (set as 100% on y‐axis). Media was collected after 2‐hr incubation and centrifuged at 2,700 g for 15 min at 4°C. Supernatant was collected and the number of extracellular vesicles (EVs) was quantified using <t>ZetaView</t> <t>Nanoparticle</t> Tracker. The data points correspond to per cent inhibition of EV release (±SEM) over a concentration range for each compound; 100% release corresponds to 4.87 × 107 ± 0.098 EVs. Results are the average of three or four independent assays. Each independent assay was the average of three replicates. pEC50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] against response, using GraphPad Prism 7
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Inhibition of EV secretion by PDDC and 5 in glial cells. Primary astrocytes were treated in parallel incubations with PDDC, 5, or cambinol in the 0.03–30 μM range as indicated; DMSO (0.02%) was used as vehicle control (set as 100% on y‐axis). Media was collected after 2‐hr incubation and centrifuged at 2,700 g for 15 min at 4°C. Supernatant was collected and the number of extracellular vesicles (EVs) was quantified using ZetaView Nanoparticle Tracker. The data points correspond to per cent inhibition of EV release (±SEM) over a concentration range for each compound; 100% release corresponds to 4.87 × 107 ± 0.098 EVs. Results are the average of three or four independent assays. Each independent assay was the average of three replicates. pEC50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] against response, using GraphPad Prism 7

Journal: British Journal of Pharmacology

Article Title: A novel and potent brain penetrant inhibitor of extracellular vesicle release

doi: 10.1111/bph.14789

Figure Lengend Snippet: Inhibition of EV secretion by PDDC and 5 in glial cells. Primary astrocytes were treated in parallel incubations with PDDC, 5, or cambinol in the 0.03–30 μM range as indicated; DMSO (0.02%) was used as vehicle control (set as 100% on y‐axis). Media was collected after 2‐hr incubation and centrifuged at 2,700 g for 15 min at 4°C. Supernatant was collected and the number of extracellular vesicles (EVs) was quantified using ZetaView Nanoparticle Tracker. The data points correspond to per cent inhibition of EV release (±SEM) over a concentration range for each compound; 100% release corresponds to 4.87 × 107 ± 0.098 EVs. Results are the average of three or four independent assays. Each independent assay was the average of three replicates. pEC50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] against response, using GraphPad Prism 7

Article Snippet: Extra cellular vesicles were quantified using ZetaView Nanoparticle tracker (Particle Metrix GmBH, Meerbusch, Germany) and the corresponding Zetaview software (8.03.04.01). pEC 50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] vs. response using GraphPad Prism 7.

Techniques: Inhibition, Incubation, Concentration Assay

Inhibition of EV secretion by PDDC and 5 in glial cells. Primary astrocytes were treated in parallel incubations with PDDC, 5, or cambinol in the 0.03–30 μM range as indicated; DMSO (0.02%) was used as vehicle control (set as 100% on y‐axis). Media was collected after 2‐hr incubation and centrifuged at 2,700 g for 15 min at 4°C. Supernatant was collected and the number of extracellular vesicles (EVs) was quantified using ZetaView Nanoparticle Tracker. The data points correspond to per cent inhibition of EV release (±SEM) over a concentration range for each compound; 100% release corresponds to 4.87 × 107 ± 0.098 EVs. Results are the average of three or four independent assays. Each independent assay was the average of three replicates. pEC50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] against response, using GraphPad Prism 7

Journal: British Journal of Pharmacology

Article Title: A novel and potent brain penetrant inhibitor of extracellular vesicle release

doi: 10.1111/bph.14789

Figure Lengend Snippet: Inhibition of EV secretion by PDDC and 5 in glial cells. Primary astrocytes were treated in parallel incubations with PDDC, 5, or cambinol in the 0.03–30 μM range as indicated; DMSO (0.02%) was used as vehicle control (set as 100% on y‐axis). Media was collected after 2‐hr incubation and centrifuged at 2,700 g for 15 min at 4°C. Supernatant was collected and the number of extracellular vesicles (EVs) was quantified using ZetaView Nanoparticle Tracker. The data points correspond to per cent inhibition of EV release (±SEM) over a concentration range for each compound; 100% release corresponds to 4.87 × 107 ± 0.098 EVs. Results are the average of three or four independent assays. Each independent assay was the average of three replicates. pEC50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] against response, using GraphPad Prism 7

Article Snippet: Extra cellular vesicles were quantified using ZetaView Nanoparticle tracker (Particle Metrix GmBH, Meerbusch, Germany) and the corresponding Zetaview software (8.03.04.01). pEC 50 values and corresponding 95% confidence asymmetrical intervals, CI (95%), were obtained from a non‐linear fit of log [inhibitor] vs. response using GraphPad Prism 7.

Techniques: Inhibition, Incubation, Concentration Assay