| Autor: |
Marinelli, Carla, Liddo, Rosa Di, Facci, Laura, Bertalot, Thomas, Conconi, Maria, Zusso, Morena, Skaper, Stephen, Giusti, Pietro |
| Rok vydání: |
2015 |
| DOI: |
10.6084/m9.figshare.c.3598415_d1 |
| Popis: |
Figure S1–S4. Figure S1 l-leucyl-l-leucine methyl ester (L-LME) depletes enriched astrocytes of microglia. Following mechanical separation of microglia from the mixed glial cell monolayers, astrocytes were detached, replated, and incubated the next day with 50 mM L-LME for 60 min. Cells were returned to fresh culture medium and processed 24 h later for Iba1 mRNA by RT-PCR. L-LME- treated astrocytes displayed an Iba1 mRNA level of 0.17-fold difference compared to enriched (≥95 %) astrocytes. Parallel sets of cultures were processed for GFAP (red) and Iba1 (green) immunocytochemistry (lower panels). Nuclei are labelled blue with DAPI. Note the loss of residual microglia (arrows) in the L-LME-treated culture. Figure S2 The NF-κB inhibitor Ro-106-9920 blocks LPS-induced TNF-α gene and protein expression in purified rat cortical microglia. Cells were pretreated 30 min with 1 μM Ro-106-9920 (‘Ro-106’), followed by addition of LPS (100 ng/ml final) and incubation continued for a further 6 h. Cells were then collected and processed for mRNA analysis by RT-PCR (left panel) and culture medium for TNF-α content by ELISA (right panel). Values are means ± s.e.m. (n = 3). ***p |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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