Fig. 2
PIXR is induced upon feeding in I. scapularis nymphs and larvae. Quantitative RT-PCR assessment of pixr transcript levels in: a unfed clean (UF clean gut) and unfed B. burgdorferi-infected (UF Bb gut) gut, fed salivary glands from clean (Fed clean SG) and B. burgdorferi-infected nymphs salivary glands (Fed Bb SG), fed guts of clean (Fed clean guts) and Borrelia-infected (Fed Bb gut) nymphs. Each data point represents a pool of three nymphal salivary glands or guts. b clean unfed larvae (Unfed larvae) and larvae fed on clean (Fed clean larvae) or B. burgdorferi-infected mice (Fed Bb larvae). Each data point represents a pool of five fed larvae; Horizontal bar represents the median. Mean values significantly different in a non-parametric Mann–Whitney test (P < 0.05) indicated. c Coomassie blue staining of purified recombinant PIXR (rPIXR) produced in a Drosophila expression system, lane 1. Protein markers shown to the left (Precision Plus Biorad protein markers). d Western blot of protein extracts of fed clean nymphal salivary glands (SG) and gut (Gut) probed with polyclonal rabbit anti rSalp14 sera, anti-rPIXR or anti rSalp25D sera. e Replicate western blots of protein extracts of unfed larvae lane 1; unfed B. burgdorferi-infected nymphal gut, lane 2; unfed clean nymphal gut, lane 3; larvae fed on B. burgdorferi-infected, lane 4; or on clean mice, lane 5; guts of fed B. burgdorferi-infected nymphs lane 6; or fed clean nymphs lane 7; probed with polyclonal naive rabbit sera, or polyclonal anti-rPIXR or anti rSalp25D rabbit sera. Protein markers (Precision Plus Biorad protein markers) are shown in lane M and their cognate molecular weights denoted to the left. Arrows indicate PIXR and Salp25D-specific protein bands
