Table 1.
Assay optimization parameters.
Table 2.
Primers used in this study.
Fig 1.
Baseline corrected fluorescence values (mean ± SE) obtained by incubating resazurin with D. immitis microfilariae of two different strains (JYD-34 and Big Head) at four different concentrations (50 to 500 microfilaria per well) for incubation periods ranging from 10 minutes to 24 hours at 37°C.
Panel A: Values arranged by incubation time. Panel B. Values arranged by microfilariae concentration. Correlation values and p-values are shown in panel B.
Fig 2.
Baseline corrected fluorescence values (mean ± SE) obtained by incubating Hoechst 33342 with D. immitis microfilariae of two different strains (JYD-34 and Big Head) at four different concentrations (50 to 500 microfilaria per well) for incubation periods ranging from 10 minutes to 24 hours at 37°C.
Panel A: Values arranged by incubation time. Panel B. Values arranged by microfilariae concentration. Correlation values and p-values are shown in panel B.
Fig 3.
Baseline corrected absorbance values of resorufin (panels A and B) and resazurin (panels C and D) (mean ± SE) obtained by incubating resazurin with D. immitis microfilariae of two different strains (JYD-34 and Big Head) at four different concentrations (50 to 500 microfilaria per well) for incubation periods ranging from 10 minutes to 24 hours at 37°C. Panel A and C: Values arranged by incubation time. Panel B and D. Values arranged by microfilariae concentration. Correlation values and p-values are shown in panel B.
Fig 4.
Baseline corrected fluorescence values (mean ± SE) obtained by incubating Resazurin (Panel A) and Hoechst 33342 (Panel B) with D. immitis microfilariae of five strains (Berkeley, Big Head, GA3, JYD34 and LOL) at four different concentrations (50 to 500 microfilaria per well) for 1 hour at 37°C. Due to low microfilaremia, 500 microfilaria per well could not be tested for Berkley. The coefficient of determination for the linear models (R2) is shown.
Fig 5.
Baseline corrected fluorescence values (mean ± SE) obtained by incubating drugs with 300 D. immitis microfilariae of 5 strains for 1 hour, followed by incubation with resazurin (Panel A) or Hoechst 33342 (Panel B) for 1 hour. Two-way ANOVA with Tukey’s multiple comparison was performed. Only comparisons between strains for each treatment are shown.
Fig 6.
Relative expression of P-glycoprotein and cytochrome P450s measured using qPCR.
Two-way ANOVA with false discovery rate correction was performed. q-values are reported.