Fig. 3: Cellular uptake.

Following a 10 µM (**P = 0.0035, **P = 0.0029, ****P < 0.0001 NM1TFV vs. NM2TFV prodrug; ****P < 0.0001 NM1TFV vs. TAF TFV-DP) or b 100 µM treatments (**P = 0.0021, ***P = 0.0003 NM1TFV vs. NM2TFV prodrug; *P = 0.0273, **P = 0.0022 NM1TFV vs. NM2TFV TFV-DP), intracellular prodrug (left) and TFV-DP (right) concentrations in MDM was measured over an 8 h period. a, b Prodrug uptake of NM1TFV (green) significantly greater than NM2TFV (orange) and TAF (blue). TAF treatment resulted in the most rapid prodrug conversion to TFV-DP, followed by NM2TFV, and NM1TFV. a, b Values reported are the mean ± SEM of three replicates. Each experiment was repeated independently three times with equivalent results. c Transmission electron microscopy (TEM) of control, TAF, NM1TFV, and NM2TFV loaded MDM after 8 h drug incubation with 10 µM drug. Scale bar: 2 μm. For comparison of two groups Student’s t-test (two-tailed, unpaired) was used (*P ≤ 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 NM1TFV compared with NM2TFV). A one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test was used to compare the prodrug and TFV-DP levels among three treatment groups (*P ≤ 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 NM1TFV compared with NM2TFV or TAF). Data were independently reproduced three times.