Fig. 4

ALS-CSF regulates the synthesis and release of the cytokines. The analysis of mRNA expression uponALS-CSF exposure revealed an upregulation of IL-6 (a *p < 0.05, NC; ##p < 0.01, NALS vs. ALS; n = 5 in triplicates), TNF-α (b **p < 0.01, NC and NALS vs. ALS; n = 5 in triplicates), and IL-10 (d ***p < 0.001, NC and NALS vs. ALS; n = 5 in triplicates) mRNA. However, the mRNA expression of IFN-γ was downregulated (c ***p < 0.001, NC and NALS vs. ALS; n = 5 in triplicates). This figure also shows representative confocal images of microglial cultures immunolabeled for IL-6 (e–e”), TNF-α (f–f”), IFN-γ (g–g”), and IL-10 (h–h”) in the control (NC), NALS, and ALS-CSF groups. Note the prominent perinuclear localization of cytokines and multinucleation in the microglia of the ALS group. The histogram represents the mean fluorescence intensity of cytokines in all the study groups. Note the increased levels of IL-6 (i *p < 0.05, NC; ##p < 0.01, NALS vs. ALS; n = 5 in duplicates) and TNF-α (j **p < 0.01, NC and NALS vs. ALS; n = 5 in duplicates) in the ALS group compared to the control groups. IL-10 was downregulated (l **p < 0.01, NC and NALS vs. ALS; n = 5 in duplicates), while IFN-γ levels (k) remain unchanged. Statistical significance was assessed using oneway ANOVA followed by Tukey’s post hoc test