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. 2017 Jun 19;159(1):64–75. doi: 10.1093/toxsci/kfx117

Figure 5.

Figure 5

Astrocytes are a cellular origin of rotenone-induced Cxcl1. Total mRNA was collected from primary mixed glial cultures reverse transcribed and analyzed using Real-Time PCR. Rotenone treatment significantly elevated Cxcl1 (A) and IL1b (B) mRNA levels as compared with LPS treatment alone (Representative experiment conducted in triplicate, n = 3 per group, *p value < .05 Ordinary 1-way ANOVA with Dunnet’s Multiple Comparison Test). C, Mixed glial cultures were separated using the shake off method and separation of microglia from astrocytes was confirmed by flow cytometry anti-CD11b-FITC antibodies. The expression of Cxcl1 and IL1B mRNA was assayed in purified astrocytes and microglia using Real-Time PCR. Rotenone treatment significantly elevated Cxcl1 (D) and IL1b (E) mRNA levels as compared with LPS treatment alone. A significant majority of Cxcl1 expression was detected in CD11b-negative astrocytes (D) while a significant majority of IL1b (E) expression was detected in CD11b-positive microglia (Representative experiment conducted in triplicate, n = 3 per group, **p value < .01 Ordinary 1-way ANOVA with Dunnet’s Multiple Comparison Test).