Fig 2: Western blot analysis. The expression of iNOS in the ipsilateral and contralateral DRGs 4 h, 24 h, and 7 days after sciatic-nerve transection. (a) Nuclear fraction; (b) cytoplasmic fraction. One-way ANOVA. Mean ± SEM. n = 6. * p < 0.05, ** p < 0.01, *** p < 0.001—ipsilateral DRG with respect to the contralateral DRG of the same animal; # p < 0.05, ### p < 0.001—ipsilateral DRG with respect to the ipsilateral DRG, at different time intervals (with respect to 4 h and 24 h).

Fig 3: Mycobacterial WhiB3 and DosR depletion compromises bone morphogenesis protein (BMP) signaling and associated immune responses. Peritoneal macrophages from C57BL/6 mice were infected with mentioned strains of Mycobacterium tuberculosis (Mtb) [multiplicity of infection (MOI) 1:10] for 12 h in each of the following cases (A–D). Hallmarks of activated BMP signaling and iNOS were analyzed by immunoblotting (A). Transcript levels of miR27a were assessed by RT-PCR (B). Mouse peritoneal macrophages were pretreated with Poly I:C (1 h) and infected with mentioned strains of Mtb as indicated. Total protein levels of TICAM1 were assessed by immunoblotting (C). Transcript levels of Ifn-a and Ifn-ß were assessed by quantitative real-time RT-PCR (D). Model: Mtb result not only in the activation of cellular c-Abl but also in its nuclear translocation. In the nucleus, c-Abl associates with acetyltransferase KAT5 and recruits the transcription factor TWIST1 to BMP2/4 promoters resulting in its activation. Activated BMP signaling keeps Mtb stimulated host protective iNOS levels in check. It is also accompanied by a surge in the levels of miR27a that is a negative regulator of TICAM1, thus stifling TLR3-dependent innate responses (E). All in vitro Mtb infections were performed for 12 h, and data represent the mean ± SEM for five values from three independent experiments, **P < 0.005, ***P < 0.0001 (one-way ANOVA followed by Tukey’s multiple-comparisons test) and all blots are representative of three independent experiments.

Fig 4: Western blot analysis. The effect of the NO donor (SNP) and the iNOS selective inhibitor (SMT) of p53 expression in the ipsilateral and contralateral DRGs, 4 h and 24 h after sciatic-nerve transection. (a) Nuclear fraction; (b) cytoplasmic fraction. One-way ANOVA. Mean ± SEM. n = 6. * p < 0.05, ** p < 0.01—ipsilateral DRG with respect to the contralateral DRG of the same animal; # p < 0.05, ## p < 0.01—ipsilateral DRG with respect to the ipsilateral DRG of the control group animal, within the framework of the same time interval after axotomy.

Fig 5: Bone morphogenesis protein (BMP) signaling regulates iNOS, aiding evasion of host immune system during mycobacterial infection. Mouse peritoneal macrophages were pretreated with Imatinib/Compound C/DPI/1400W followed by Mycobacterium tuberculosis (Mtb) infection where indicated. Reactive oxygen species production was estimated by dichlorofluorescin diacetate (DCFDA) fluorescence (A) and levels of nitrite were assessed by Griess assay (B). iNOS was assessed by immunoblotting in mouse peritoneal macrophages pretreated with Imatinib or Compound C, followed by Mtb infection [(C), left] and in RAW264.7 macrophages transiently transfected with Abl1 siRNA subsequently followed by Mtb infection [(C), right]. Phosphorylated STAT1 and STAT3 were assessed by immunoblotting in each of the following instances (D–F). RAW264.7 macrophages were transiently transfected with pcDNA3.1 or BMPR2 DN or BMPR1a CA construct followed by pretreatment with IFN-? (200 U/ml, 1 h) and subsequent Mtb infection as indicated (D). RAW264.7 macrophages were transiently transfected with pcDNA3.1/c-Abl OE/c-Abl kinase dead (KD) construct followed by pre-treatment with IFN-? (200 U/ml, 1 h) and subsequent Mtb infection as indicated (E). RAW264.7 macrophages were transiently transfected with pcDNA3.1/c-Abl OE/c-Abl KD construct followed by pretreatment with Poly I:C (1 h) and subsequent Mtb infection as indicated (F). All in vitro Mtb infections were performed for 12 h, and data represent the mean ± SEM for five values from three independent experiments. ns, not significant; *P < 0.05, **P < 0.005, ***P < 0.0001 (one-way ANOVA followed by Tukey’s multiple-comparisons test) and all blots are representative of three independent experiments.