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Fig. 3 | Journal of Neuroinflammation

Fig. 3

From: HIV-tat alters Connexin43 expression and trafficking in human astrocytes: role in NeuroAIDS

Fig. 3

HIV-tat did not increase Cx30 and mRNA protein expression and did not contribute to the enhanced gap junctional communication induced by HIV-tat in human primary astrocytes. Human primary astrocytes were treated with recombinant HIV-tat protein (100 ng/ml), and expression and function of Cx30-containing channels were analyzed. a Labeling for nuclei (DAPI, blue staining), Cx30 (Alexa 488, green staining), and GFAP (Cy3, red staining) in untreated conditions (control) and after 24 h HIV-tat treatment. The last panel represents the merge of all colors. Bar 75 μm. b qRT-PCR for Cx30 and GAPDH using untreated and HIV-tat-treated (24 h) cultures of astrocytes. No significant differences in mRNA Cx30 expression were detected in control cells (white bars). HIV-tat treatment decreased Cx30 mRNA expression in a time-dependent manner (# p ≤ 0.001, n = 4, black bars). c Western blot analysis of Cx30 protein expression in control and HIV-tat-treated human astrocytes. Tubulin (tub) was used as a loading control. As a Cx30 positive control, mouse brain was used (+). HIV-tat decreased Cx30 expression in a time-dependent manner (Fig. 2c). d To examine the contribution of Cx30 to the increased gap junctional communication induced by HIV-tat, we reduced further the expression of Cx30 using siRNA. In this condition, we reduced Cx30 by at least 80 %; however, no changes in dye coupling were observed, suggesting that Cx43 mediate most of the communication (n = 4, p ≤ 0.005)

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