Western blot analyses for 2′ , 3′-cyclic nucleotide 3′-phosphodiesterase (CNPase). a. Western blots for CNPase. Ten-lane precast gels were used. Each lane had an individual 5 μl sample of CSF from the categories of patients shown. After protein separation using SDS PAGE they were blotted onto a PVDF membrane for probing with antibodies. ECL detection was used to visualise the labelled bands and for semi-quantitative analysis using Image-J software. A single 45 KDa band was labelled using the antibody to CNPase and was detected in all samples as shown in these representative gels. All samples were run a minimum of three times with a total number of samples as shown for semi-quantitative analysis. Patient groups as for Figure 1. b. Semi-quantitative analysis of the 45 KDa band labelled with anti-CNPase antibody shows similar levels of CNPase to that in normal CSF in FOH, PHH and SB/HC but in the LOH group where a significant reduction (P < 0.05 indicated by asterisk *) was observed compared to normal and other hydrocephalic groups. In the LOH gel 3 of the 4 samples show reduced CNPase compared to normal and one sample not. Increased numbers will provide better resolution of this particular group. Values are Mean ± SEM of Normal n = 4, FOH n = 4, LOH n = 4, PHH n = 4, SB/HC n = 4.