Anti cd68

We digested and homogenized regenerated sciatic nerves that were homogenized in lysis buffer according to standard procedure. In brief, 20 μg protein was loaded on 10% sodium dodecyl sulphate polyacrylamide gel for electrophoresis and was immediately transferred to nitrocellulose sheets for 3 hours. All the samples were transferred to a poly (vinylidene fluoride) membrane and incubated at 4°C overnight. The primary antibodies (Thermo Fisher Scientific, Carlsbad, CA, USA) were anti‐IL‐6 (1:3000), anti‐TNF‐α (1:2500) and anti‐CD68 (1:1500). β‐Actin was used as the control. The relative quantitative analysis was dependent on ImageJ evaluation of the blot's grey values.

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).

The immunoblot analysis was performed as described previously 35 (link). Briefly, tissues and cells were lysed in 1×RIPA buffer (Sigma-Aldrich, #R0278). Equal amounts of protein in each sample were loaded into a 10% SDS-PAGE gel. After transferring to a membrane and blocking with 5% milk, the proteins were probed with the following primary antibodies: anti-CtBP1 (BD Biosciences, San Jose, CA, USA, #612042), anti-CtBP2 (BD Biosciences, #612044), anti-CD31 (ThermoFisher Scientific, #PA5-16301), anti-CD55 (ThermoFisher Scientific, #PA5-82005), anti-CD68 (ThermoFisher Scientific, #MA5-13324), anti-GAPDH (Santa Cruz Biotechnology, Dallas, Texas, USA, #sc-365062), anti-Caspase-1 (Santa Cruz Biotechnology, #sc-56036), anti-Flag (Sigma-Aldrich, #SAB4200071), anti-Myc (Abcam, Cambridge, MA, USA, #ab9106), anti-p300 (Santa Cruz Biotechnology, #sc-585), anti-c-Jun (Sigma-Aldrich, #SAB4501606), anti-c-FOS (Sigma-Aldrich, #F7799), anti-p50 (ThermoFisher Scientific, #PA1-30409), anti-p65 (ThermoFisher Scientific, #14-6731-81), anti-IRF2 (Abcam, #ab3388), anti-STAT4 (Abcam, #ab68156), anti-NLRP3 (Abcam, #ab210491), anti-Il-1β (Abcam, #ab2105), anti-DNMT1 (Abcam, #ab13537), and anti-DNMT3A (Abcam, #ab2850). After probing with secondary antibodies, protein band signals were detected using a Pierce^TM ECL western blotting substrate (ThermoFisher Scientific, #32106).