Microglia were transfected with siRNA targeting Kv1.3 (Kv1.3-siRNA) or nonspecific GAPD control siRNA (Ctrl-siRNAS) for 48 or 72 hr, followed by an additional 24 hr exposure to Tat (200 ng/ml). Cells were then harvested for detections of Kv1.3 mRNA (48 hr post-transfection/24 hr Tat treatment) and Kv1.3 proteins (72 hr post-transfection/24 hr Tat treatment). Supernatants were subjected to neuronal culture. Neuronal apoptosis and viability assay were determined using TUNEL staining and MTT assay. A: Representative gels show RT-PCR products for Kv1.3 mRNA and internal control β-actin and bar graph reflects the density of each band after normalization of its β-actin. B: Western blots show Kv1.3 protein and internal control β-actin protein expression of microglia, and bar graph shows densitometric quantification of each band. C: Collected supernatants were subjected to primary neuronal culture at a dilution of 1:5 for 24 hr and neuronal viability was evaluated by MTT assay. An increased viability was observed in neurons treated with supernatants recovered from microglia transfected with Kv1.3-siRNA, but not transfected with Ctrl-siRNA. D: Transfection of microglia with Kv1.3-siRNA significantly reduced neuronal apoptosis. In contrast, transfection of microglia with Ctrl-siRNA exhibited no significant protective effect. E: Apoptotic neurons were visualized by fluorescence microscopy at ×400 original magnification. Scale bar equals 100 µm. * p<0.05, *** p<0.001 vs Ctrl-siRNA; ### p<0.001 vs Ctrl (blank).
Citation: The PLOS ONE Staff (2014) Correction: HIV-1 Tat Protein Increases Microglial Outward K+ Current and Resultant Neurotoxic Activity. PLoS ONE 9(9): e109218. https://doi.org/10.1371/journal.pone.0109218
Published: September 19, 2014
Copyright: © 2014 The PLOS ONE Staff. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.