Fig. 1

HIV-1 infection and HAND pathology dysregulate Ng expression in human FC tissue: a FC sections from uninfected control, HAND (−), and HAND (+) subjects were IHC-stained for Ng and counterstained with hematoxylin. Scale bar indicates 50 μm. b, c The area and intensity of Ng-positive cells were counted from five different fields from each subject and divided by the total number of cells in those fields to calculate the mean area and mean intensity per cell. Positive cells were masked based on signal intensity threshold and normalized over noise using the NIS Elements software. Selection of an appropriate background and shading correction as well as smoothing filter object minimized noise of the images, allowing for more accurate analyses. d FC tissues from control, HAND (−), and HAND (+) subjects were homogenized and lysed, and 5 μg of each lysate were loaded into each lane to measure Ng expression by western blot using specific antibody (N = 4 for each group). MAP2 was used as control for normalization. Tubulin was used as a loading control. e Band intensities were quantitated by the ImageJ software, and the data were normalized with MAP2 and tubulin. f qRT-PCR was performed with Ng-specific primers and probes using the RNA isolated from the frozen tissues from the same subjects (N = 4 for each group). Results were normalized with housekeeping gene RPLPO. Results are the ±SEM of four individual experiments, nonparametric Mann-Whitney tests were performed to calculate significance, and *p< 0.05 and **p < 0.01 compared to control. NS not significant