Over-Expression of Cysteine Leucine Rich Protein Is Related to SAG Resistance in Clinical Isolates of Leishmania donovani
Fig 1
Molecular characterization of L. donovani CLrP.
(A) Western analysis of membrane and soluble fraction of L.donovani with anti-CLrP antibody Lane1:Molecular mass marker, Lane2: Ld Membrane fraction, Lane3: Ld Soluble fraction, (A1) Lane M: Membrane fraction analyzed with anti-GRP78 antibody, Lane S: Soluble fraction analyzed with anti-GRP78 antibody (B) Immunolocalization of LdCLrP in Ld. B1-B4: Images of permeablized Ld with anti rLdCLrP and then FITC conjugated secondary antibody (arrows shows the nucleus localization of CLrP), (B5-B8) Images of non-permeabilized Ld with anti-rLdCLrP (B5) Phase contrast image (B6) Fluorescent image (rabbit raised anti-CLrP + FITC conjugated rabbit sec antibody) (B7) Fluorescent image (Rhodamine tagged Con A) and (B8) mearged image of B6 and B7. (C-D) ChIP assay probed with Ld genes through PCR (C) 60sRL23a gene (D) TPI gene M:1kb DNA ladder, Lane 1: Chromatin material obtained after immune-precipitation with anti rCLrP, Lane 2: Chromatin material obtained after immune-precipitation with anti-actin, Lane 3: Chromatin material obtained after immune-precipitation with pre-immune serum, Lane 4: Chromatin material obtained after immune-precipitation with anti GRP78. (E) Deglycosylation of WCL of Ld, Lane1: WCL of Ld Lane 2: control, Lane3: Ld+deglycosylase mix.