Balb cbyj female mice

All chemical reagents
were purchased from
Sigma–Aldrich, while anhydrous solvents and phosphate-buffered
saline (PBS, 0.01 M) were purchased from Scharlab S.L. and used without
further purification. Palbociclib was purchased from Selleckchem,
and Dulbecco’s modified Eagle medium (DMEM) and fetal bovine
serum (FBS) were purchased from Gibco. Flat-bottom clear 96-well plates
were purchased from Promega. High-resolution mass spectrometry (HRMS)
and the data was recorded with a TripleTOF T5600 (ABSciex, U.S.A.)
spectrometer. ¹H and ¹³C NMR spectra were collected
on a Bruker FT-NMR Avance 400 (Ettlingen, Germany) spectrometer at
300 K, using TMS as an internal standard. HPLC measures were obtained
by a Waters 1525 binary HPLC pump, and spectra were recorded by a
Waters 2998 photodiode array at 260 nm. Fluorescence spectra were
recorded by a JASCO FP-8500 fluorescence spectrophotometer, Luminescence
was collected in a VICTOR multilabel plate reader (PerkinElmer). Confocal
fluorescence images were taken on a Leica TCS SP8 AOBS, and two-photon
images were acquired using a multiphoton Olympus FV1000MPE confocal
microscope. Images were analyzed using ImageJ software. The SK-Mel-103
(human melanoma) cancer cell line and 4 T1 (breast cancer cells) were
acquired from the American Type Culture Collection (ATCC). BALB/cByJ
female mice were purchased from Charles River laboratories, France.

Nine-week-old BALB/cByj female mice (Charles River Laboratories) were housed in a temperature-, humidity- and light-controlled environment and had ad libitum access to tap water and breeder chow 5080 (containing 11% fat and 21% protein, Labdiet, St. Louis, MO). Following at least one week period of acclimatization, mice were mated and began the protocol as summarized in Figure 1. Mice were allowed to wean until PND28.
Animal experiments were carried out in strict accordance with the principles and procedures of the Guide for the Care and Use of Laboratory Animals and institutional guidelines. The protocol was approved by the Institutional Animal Care and Use Committee, Columbia University Medical Center.

Female BALB/cByJ mice, aged 5 to 6 weeks, were purchased from Charles River Laboratories (France). Mice were infected with a low dose aerosol (100 to 200 CFU/lung) of logarithmic-phase M. tuberculosis H37Rv bacilli and then were allocated to experimental groups and returned to their cages. Five mice were used per time point for each regimen. Treatment was initiated 4 weeks after infection.
Macozinone (MCZ), 11626091, and INH were prepared weekly in 0.5% methylcellulose and administered at 25, 50, and 10 mg/kg, respectively, by gavage 5 days a week for 4 weeks. In vivo efficacy of each treatment was assessed by CFU enumeration after plating dilutions of the lung and spleen homogenates on 7H10 agar plates containing 10% oleic acid-albumin-dextrose-catalase (OADC), cycloheximide (10 μg/ml), and ampicillin (50 μg/ml). Plates were incubated for 4 weeks at 37°C before CFU were enumerated. CFU counts were log₁₀ transformed before analysis as the mean log₁₀ CFU ± standard deviation and compared by a Student t test using GraphPad Prism version 7.0 software. P values less than 0.05 were considered statistically significant.
Experiments were approved by the Swiss Cantonal Veterinary Authority (authorization 3082) and performed between June and August 2017.

Female BALB/c By J mice were obtained from Charles River (France). The protocol was approved by the Ethics Committee on Animals Experimentation Pays de Loire (accreditation number 3455). The mice were housed in ventilated cages in the IRS-1 Experimental Therapeutics Unit with free access to water and a standard diet (SAFE A04) and a dark/light cycle of 12:12 h. The total extract of HDMs was obtained from Stallergenes Greer (Antony, France).

Female BALB/cByJ mice (6–7 weeks; Charles River, NL and Harlan, Bicester, UK) were used. All animal experiments of this study were approved by the Animal Experiments Committee of the Leiden University Medical Center (DEC 13132 and 14307). The Dutch Experiments on Animal Act is established under European guidelines (EU directive no. 86/609/EEC regarding the Protection of Animals used for Experimental and Other Scientific Purposes). All experiments were performed in accordance with relevant guidelines and regulations. Two P. yoelii (Py) lines were used: (i) the reference “wild type” Py17XNL parasite line 1971cl1 (PyWT; PyGFP-luc_con; line RMgm-689; www.pberghei.eu (Lin et al., 2011 (link)); which contains the fusion gene gfp-luc gene under control of the constitutive eef1α promoter integrated into the silent 230p gene locus (PY17X_0306600) and does not contain a drug-selectable marker and (ii) the “genetically attenuated parasite” Py17XNL mutant that lacks the gene fabb/f (3-oxoacyl-acyl-carrier protein synthase; PY17X_1126500). This mutant (ΔPyFabBF-GFP-Luc_con; PyΔfabb/f ; mutant RMgm-4109; www.pberghei.eu) was generated in the reference line 1971cl1 (Haeberlein et al., 2017 (link)) by standard methods of transfection using a DNA construct that targets the fabb/f gene containing hdhfr/fcu selectable marker cassette by double cross-over integration.