Screening of cytotoxicity of isolated compounds and checkboard assays for evaluation of potential antimycobacterial synergism activity of compound 2 (5_312) with commercially available antimicrobial agents

Screening of cytotoxicity (Dr. Timbilla Abdul Aziz, Assoc. Prof. Radim Havelek): The raw data represent WST absorbance measurements. Slots for substances in the manuscript were 37 to 42, which correspond to molecules 5/311 to 5/316 across the 3 individual replicates. Within the article, the compound designations correspond as follows: 5/311 – compound 1; 5/312 – compound 2; 5/313 – compound 3; 5/314 – compound 4; 5/315 – compound 5; 5/316 – compound 6. The remaining positions belonged to other samples. Additionally, BLANK (standard blank) and DMSO (which serves as a negative control, 0.1% DMSO) were included. Within the 3rd replicate, Jurkat and MOLT-4 cells are saved in separate files. Checkerboard assays (Dr. Ondřej Janďourek, Dr. Klára Konečná): Interaction measurements were determined by checkerboard assay. Microdilution broth method was used as tool for screening mutual interaction of compound 2 (within raw data signed as 5/312) and commercially available antimicrobial agents. Assay was performed in standard 96-well microtitration plates (Gamedium, Czech Republic). First row and column were used for MIC determination of tested compounds alone, whereas the first well in the plate was consisted of broth+DMSO and microbes only (positive growth control) and last well in the plate was consisted of broth+DMSO only (negative growth control). INH, RIF, CIP, bedaquiline (BDQ) and ethambutol (EMB) (Merck, Germany) were chosen for combinations with 5/312 (compound 2). Strains selected for determination were M. kansasii (ATCC 12478) and M. tuberculosis H37Ra (ATCC 9431) and cultivation medium was enriched Middlebrook 7H9 broth. The raw data were measured as fluorescence.