Western blot analyses for glial fibrillary acidic protein (GFAP). a. Western blots for GFAP. 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 53 KDa band was labelled using the antibody to GFAP and only found in PHH and SB/HC samples as shown in this representative gel. All samples were run on at least 3 gels with a total number in each group as noted for the semi-quantitative analysis. FOH: fetal-onset hydrocephalus, LOH: late-onset hydrocephalus, PHH: post-haemorrhagic hydrocephalus, SB/HC: spina bifida with hydrocephalus. b. Semi-quantitative analysis of the 53 KDa band labelled with anti-GFAP antibody. Values are Mean ± SEM of Normal n = 8, FOH n = 4, LOH n = 4, PHH n = 5, SB/HC n = 4. Only PHH showed a significant difference from normal which had undetectable levels so that the difference was effectively from zero protein (P < 0.05 indicated by asterisk *). SB/HC also shows an increase but this was not significantly different from normal. Rat (postnatal P1) astrocytes cell lysates were used as a positive control for the GFAP antibody and these are shown in the last lane.