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Figure 2 | Journal of Neuroinflammation

Figure 2

From: Adaptive Müller cell responses to microglial activation mediate neuroprotection and coordinate inflammation in the retina

Figure 2

Influence of microglia on Müller cell gliosis, proliferation, and apoptosis. (A) The influence of microglia on Müller cell gliosis was assessed by evaluating mRNA expression of genes typically altered in gliosis by semi-quantitative RT-PCR: glutamine synthethase (GS), glutamate aspartate transporter (GLAST), and the intermediate filament, vimentin. Representative gel images for the PCR amplification of each mRNA species, with parallel controls from which reverse transcriptase is omitted from the amplification reaction (to confirm the absence of genomic DNA amplification), are shown (right). While the expression levels of GS in Müller cells were not statistically distinct between the different co-culture conditions, levels GLAST and vimentin, typically elevated in Müller cell gliosis, were significantly decreased following co-culture with activated microglia. These results demonstrate that changes induced by microglia co-culture differed from those associated with typical Müller cells gliosis. (B) Proliferating Müller cells in culture were marked by the incorporation of BrdU and the number of proliferating cells counted and expressed as a percentage of cells present. Co-culture with unactivated microglia induced a significant decrease in Müller cell proliferation, which was further decreased with co-culture with activated microglia. (C) Müller cells undergoing apoptosis in culture were marked with TUNEL-labeling. The percentage of apoptotic Müller cells was low and similar between all three co-culture conditions. (* indicates p < 0.05 for comparisons, one-way ANOVA with Tukey-Kramer multiple comparison test, n = 6-8 replicates from two independent experiments).

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