Fig. 7: Ellipticine acts as an inhibitor of the Erg11-Ncp1 interaction.
a Schematic of Erg11-Ncp1 inhibitor screening using MYTH. b Relative growth rates of six Erg11-Ncp1 interaction candidates measured by MYTH. c Dose-matrix assays of albicans cmp1-aidΔ/cmp1-aidΔ with ellipticine and FLC; DTT as positive control. d Growth inhibition curve assays were conducted to assess the combined effects of FLC and ellipticine. Data represent mean ± SD (n = 3). e Co-IP analysis shows ellipticine blocked the Erg11-Ncp1 interaction. “Input” is the Erg11-13Myc/3HA-Ncp1 lysate used for immunoblotting. “NC” is the wild-type SN152 lysate as a negative control. The binding rate of 3HA-Ncp1 to Erg11-13Myc was quantified. f Intracellular ROS was quantified after treatment with 16 μg/mL ellipticine and 4 μg/mL FLC. g Misfolded proteins in ERG11/erg11Δ mutants were analyzed after treatment with 16 μg/mL ellipticine and 4 μg/mL FLC. h Mitochondrial misfolded proteins were determined by MitoTracker Green. i Intracellular Ca2+ levels were quantified following treatment with 16 μg/mL ellipticine and 4 μg/mL FLC in the ERG11/erg11Δ mutant. j Mitochondrial membrane potential changes were assessed by Rhodamine 123 staining. k CYC3 gene expression was quantified after treatment with 16 μg/mL ellipticine and 4 μg/mL FLC. l Annexin V/PI double staining density plots were generated. m Quantification of Annexin V/PI double staining assay. n Ellipticine protects Ncp1 from pronase degradation in DARST assays. Uncropped, unprocessed scans with molecular weight markers are provided in the Source Data file. The quantification shows 3HA-Ncp1 protein levels in the control group relative to the ellipticine group. The representative images shown are from 3 biological repeats (e, I, n). Data were presented as mean ± SD of n = 3 biological replicates (b, e–k, m, n). Two-tailed unpaired t-test (b, e–k, m, n). Source data are provided as a Source Data file.
