Hy 112675

HEI‐OC1 cells were cultured in DMEM (Gibco, USA) medium containing 10% FBS under room temperature (about 33°C) and 10% CO₂ to reach the density of 70%–90% and then to perform the passage experiments. In the experimental group, 4‐OI (250 μM; HY‐112675, MedChemExpress, USA) was added 3 h in advance. The cisplatin treatment was given at a concentration of 30 μM for 24 h. The Ras‐selective lethal 3 (RSL3) (HY‐100218A, MedChemExpress, USA) treatment was given at a concentration of 4 μM for 24 h.

The NTM strains used in this study are listed in Table 1. All clinical samples are from CF patients while lettered samples are mixed morphology cultures from the same date and same patient. Bacteria were cultured at 37° C in 7H9 broth with OADC (Remel, R450605). For minimal media experiments, a modified 7H9 broth was used based on the methods described by Muñoz-Elías and McKinney [58 (link)], specifically, NTM were grown in 7H9 + 0.5% albumin, 0.085% NaCl, 0.05% Tween-80, and carbon substrate (10 mM short chain fatty acid) for the indicated number of days. For the pH adjusted minimal media, the media with all the above listed components was pH adjusted and then filter sterilized prior to SCFA addition and use. IA (Sigma-Aldrich, I29204) was resuspended in sterile ddH2O, pH adjusted (for the pH = 7 IA), filter sterilized, and added to the 7H9 media to the desired concentration. For pH-neutral IA experiments, IA received NaOH to bring the pH to a neutral range before being brought to the desired final volume and filter sterilized for addition to the 7H9 media. 4-Octyl itaconate (MedChem Express, HY-112675) was resuspended in DMSO and added to THP-1 cells at the desired concentrations. The short chain fatty acids (SCFA) acetate, propionate, and butyrate (Sigma-Aldrich, S2889, P5436, & 303410, respectively) were resuspended in sterile ddH20 and filter sterilized prior to use.