Journal: bioRxiv

Article Title: Micafungin exposure drives multidrug resistance in Clavispora lusitaniae

doi: 10.64898/2026.06.29.735437

Figure Lengend Snippet: Micafungin-selected mutants have increased growth in all three antifungal classes, recapitulated by engineered deletions. Minimum inhibitory concentrations (MIC 50 ) determined by the static broth microdilution assay for ( A ) MCF (0-8 µg/mL), ( B ) FLC (0-256 µg/mL), and ( C ) by E-test for AMB (0-32 µg/mL). All MIC values available in Table S3. Replicates are indicated by gray points. ( D-F ) 96-well growth curve analysis of the in vitro evolved mutants and progenitor. Area Under the Curve (AUC) measured by OD 600 from growth curve assays in: ( D ) YPAD + 0.025 µg/mL MCF; ( E ) YPAD + 0.5 µg/mL FLC; or ( F ) YPAD. Replicates are indicated by gray points. Asterisks represent p < 0.05 relative to progenitor. Significant differences determined by ANOVA with Tukey HSD post-hoc analysis with adjustments for multiple comparisons (Table S3 for p-values). ( G-I ) Null mutants engineered in the RSY426 genetic background. AUC measured by OD 600 from growth assays in: ( G ) YPAD + 0.025 µg/mL MCF ( osh2Δ , p = 0.0001; erg3Δ , p = 0.0 ); ( H ) YPAD + 0.5 µg/mL FLC ( osh2Δ, p = 0.08; erg3Δ, p = 0.03 ); or ( I ) YPAD. Asterisks represent p < 0.05 relative to RSY426. Significant differences determined by ANOVA with Tukey HSD post-hoc analysis and adjustments for multiple comparisons (Table S3).

Article Snippet: Minimum inhibitory concentration (MIC) values for both FLC (ThermoFisher; 86386-73-4) and MCF (MedChemExpress; 208538-73-2) were determined using the broth microdilution method ( ).

Techniques: Microdilution Assay, In Vitro

Journal: bioRxiv

Article Title: Micafungin exposure drives multidrug resistance in Clavispora lusitaniae

doi: 10.64898/2026.06.29.735437

Figure Lengend Snippet: Area Under the Curve (AUC) measured by OD 600 from 48 hour growth curve assays in the presence and absence of the Hsp90 inhibitor, radicicol (RAD) for the C. lusitaniae progenitor, evolved ERG3 point mutants, and clinical isolate with ERG3 mutation. ( A ) AUC in YPAD in the absence (-) or presence (+) of 2.5 µg/mL RAD. ( B ) AUC in 0.025 µg/mL MCF in the absence (-) or presence (+) of 2.5 µg/mL RAD. ( C ) AUC in 0.5 µg/mL FLC in the absence (-) or presence (+) of 2.5 µg/mL RAD. Gray points indicate technical replicates. Significance (p < 0.05, ANOVA with Tukey HSD) indicated with asterisks or ‘n.s.’ for not significant (p ≥ 0.05) (Table S4). ( D, E ) Median fluorescence intensity of ( D ) Calcofluor white that stains chitin (10 µg/mL) or ( E ) hDectin that stains 1,3-β-D-glucans (15 µg/mL) measured by flow cytometry (n ≥ 10,000 cells per isolate per replicate). Asterisk represents p < 0.05 relative to progenitor determined by ANOVA with Tukey HSD post-hoc analysis and adjustments for multiple comparisons (Table S5). ( F ) Serial dilutions (10 6 -10 2 ) of evolved ERG3 mutants and progenitor isolate were spotted on agar plates with YPAD or YPAD + 15 µg/mL Calcofluor white, + 150 µg/mL Congo Red, or + 0.005% Sodium Dodecyl Sulfate (SDS). All plates were repeated in triplicate, incubated at 30°C and imaged at 24 hours.

Article Snippet: Minimum inhibitory concentration (MIC) values for both FLC (ThermoFisher; 86386-73-4) and MCF (MedChemExpress; 208538-73-2) were determined using the broth microdilution method ( ).

Techniques: Mutagenesis, Fluorescence, Flow Cytometry, Incubation

Journal: bioRxiv

Article Title: Micafungin exposure drives multidrug resistance in Clavispora lusitaniae

doi: 10.64898/2026.06.29.735437

Figure Lengend Snippet: Representative GC-MS chromatograms for the following isolates: ( A ) the drug-sensitive C. lusitaniae clinical isolate AMS5200 and progenitor for the current study ; ( B ) the in vitro evolved ERG3 A149P mutant; and ( C ) the in vitro evolved ERG3 Y304* mutant. GC-MS data obtained from cells grown for 4 hours in 0 µg/mL antifungals (No drug), 0.5 µg/mL fluconazole (FLC), or 0.016 µg/mL micafungin (MCF). Retention times indicated by values adjacent to peaks for known standards.

Article Snippet: Minimum inhibitory concentration (MIC) values for both FLC (ThermoFisher; 86386-73-4) and MCF (MedChemExpress; 208538-73-2) were determined using the broth microdilution method ( ).

Techniques: Gas Chromatography-Mass Spectrometry, In Vitro, Mutagenesis

Journal: bioRxiv

Article Title: Micafungin exposure drives multidrug resistance in Clavispora lusitaniae

doi: 10.64898/2026.06.29.735437

Figure Lengend Snippet: ERG3 mutants have a unique sterol profile consistent with Erg3 loss of function. ( A ) Mean relative abundance of sterols identified using targeted GC-MS after growth in the absence of antifungals or after growth + 0.5 µg/mL FLC. Mean value was calculated using three biological replicates per strain and per condition. See Table S6 for p-values. Asterisks indicate unidentified sterol that has similarity to Gramisterol. ( B ) Mean relative abundance of sterols identified using targeted GC-MS after growth in the absence of antifungals or after growth in 0.016 µg/mL MCF. Mean value was calculated using three biological replicates per strain and per condition. See Table S6 for p-values indicating significant differences in abundance of sterols. ( C ) Average response area of sterols without standards identified using GC-MS analysis after growth in the absence of antifungals or after growth + 0.016 µg/mL MCF or 0.5 µg/mL FLC. ( D ) Ergosterol biosynthesis pathway inferred from ( , , , ). Blue font indicates sterols synthesized as part of the alternative pathway, initiated following azole inhibition of Erg11. Biosynthetic steps where Erg3 functions are indicated in bold.

Article Snippet: Minimum inhibitory concentration (MIC) values for both FLC (ThermoFisher; 86386-73-4) and MCF (MedChemExpress; 208538-73-2) were determined using the broth microdilution method ( ).

Techniques: Gas Chromatography-Mass Spectrometry, Synthesized, Inhibition

Journal: iScience

Article Title: Universal principles of cell population growth follow from local contact inhibition

doi: 10.1016/j.isci.2026.115953

Figure Lengend Snippet: Experimental observations of cancer cells distributed at low and high density (A and B) Breast cancer cell line MCF-7 at low and high density. (C and D) Breast cancer cell line MDA-MB-231 at low and high density. The scale bars on all panels represent 50 μm in length.

Article Snippet: MCF-7 (human breast adenocarcinoma, female) , ATCC , RRID: CVCL_0031.

Techniques: