Fig. 8

Assessment of astrocytic mediated neuronal damage. a) Percentage change in the extracellular release of glutamate in transfected PDA (n = 4). b) Immunofluorescence images of neurons post 24 h of treatment with astrocytic condition media (ACM) transfected for 24 h, stained with TUNEL (red dots), and counterstained with DAPI for nuclei identification, Scale bar indicates 50 μm. c) The TUNEL positive cells were counted from five random fields from each set and divided by the total number of cell in the respective field, and fold change represents the TUNEL positive neurons treated with ACM (n=3). d) Immunofluorescence images of neurons post 24 h of treatment with astrocytic condition media of transfected PDA, neurons were labelled with anti-MAP2 antibody in red for the detection of neurite outgrowth and counterstain with DAPI for nuclei identificatiom, Scale bar is 50 μm. e) Change in the neurites length in treated neurons with ACM was determined using imageJ, and five images were randomly selected from each treatment set (n = 3). f) Fold change showing extracellular ATP release in treated neurons with ACM after 24 h of transfection with various plasmids (indicated in different colors, shown in key). Color shades represent black blocks which are for plasmid control (PC), dark grey which are HIV-1 Tat, white blocks which are overexpressed mortalin, and light grey which are cotransfectd with mortalin and HIV-1 Tat (n = 3). All data represents mean ± standard devitaion, from independent experiments (n stands for number of independent experiment), *p< 0.05, **p< 0.005 with respect to control, #p< 0.05, ##p< 0.005 with respect to HIV-1 Tat and cotransfected group (PC stands for plasmid control, and PC+PC is used as plasmid control in cotransfected cells)