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. Author manuscript; available in PMC: 2017 Aug 1.
Published in final edited form as: Glia. 2016 Jun 1;64(8):1396–1415. doi: 10.1002/glia.23013

FIGURE 2. LPS enhance the activity and the expression of Cav1.2 Ca++ channels in astrocytes.

FIGURE 2

(A) After 6DIV, astrocytes were treated with different concentrations of LPS for 3 consecutive days and were stained with antibodies against GFAP and s100β. Scale bar = 120μm. (B) The fluorescent intensity for each marker was measured by confocal microscopy and was plotted as integrated optical density (IOD). (C) Individual GFAP-positive cells were scored according to their morphological complexity in two categories. (D) After 3 days of LPS (1μg/ml) treatment, astrocytes were stained with antibodies against Cav1.2 and Cav1.3. The fluorescent intensity for each L-type Ca++ channel was measured by confocal microscopy and was plotted as IOD. Scale bar = 80μm. (E) Western blots analysis of Cav1.2/1.3 VOCC α1 subunits expression in cortical astrocytes. The analysis was performed after 3 days of LPS (1μg/ml) treatment using p84, β-actin and GAPDH as internal standards and data from three independent experiments are summarized based on the relative spot intensities. (F) After 3 days of LPS (1μg/ml) treatment, VOCC activity was examined in cultured astrocytes using fura-2 as intracellular Ca++ indicator. Note that each trace corresponds to a single cell and the horizontal bars indicate the time of addition of external solution containing high K+. (G) The bar graph shows the average amplitude of the Ca++ response, calculated from the responding cells expressed as a percentage of change of the emission intensities. Verapamil and nifedipine were applied at 5μM. Values are expressed as mean ± SEM of at least six independent experiments. *p<0.05, **p<0.01, ***p<0.001 vs. control; ###p<0.001 vs. LPS.