Validation of Microarray Findings in Lung Tissue

To validate the findings originating from microarray analysis, multilabel immuno-histochemistry was performed on formalin fixed paraffin embedded (FFPE) sections of lungs derived from RMs with ATB or LTBI, as described previously (11 (link), 30 (link)). The lung sections were stained for macrophages with anti-CD68 (Thermo Fischer Scientific, Cat no #MA5-13324) and anti-MX1 (Thermo Fischer Scientific, Cat no #PA5-22101) antibodies to validate the in-vivo expression of these markers in lung tissue. DAPI (Thermo Fischer Scientific, Cat no #D1306) was used to stain nuclei. Images were captured using Zeiss Axio ScanZ1 and Zeiss LSM-800 confocal microscope and analysis was done using Zeiss Zen 3.6 (blue edition).