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microRNA-34a-Mediated Down-Regulation of the Microglial-Enriched Triggering Receptor and Phagocytosis-Sensor TREM2 in Age-Related Macular Degeneration

Fig 3

TREM2 and DAPI nuclear staining of C8B4 murine microglial (MG) cells (A): (a) control MG cells cultured 3 days, magnification 20x; (b) treated with TNFα; (c) treated with 50 nM miRNA-34a-sc (24 hr) or (d) treated with 50 nM miRNA-34a (24 hr); note significantly reduced TREM2 protein signals in stressed MG cells (b and d) compared to control (a) or miRNA-34a-sc-treated MG cells (C); Westerns blots were performed for TREM2 using an antibody directed against the 277 amino acid murine TREM2 (M227;sc-48765) or TYROBP (DAP12; C-20; sc-7853); SCBT, Santa Cruz, California, USA); nuclei stained with DAPI as in (Fig 6); (B) (upper panel) representative Western blot and (B) (lower panel) bar graph analysis of TREM2 and DAP12 protein levels in control, TNFα-, miRNA-NC or miRNA-34a-stressed MG cells; in this sample set TREM2 protein levels were found to be significantly reduced in TNFα- or miRNA-34a-treated MG cells compared to age matched controls; there were no significant differences in the abundance of the TYROBP/DAP12 adaptor protein amongst control, TNFα, miRNA-NC or miRNA-34a treated cells (Fig 7); N = 8; *p<0.05, **p<0.01 (ANOVA).

Fig 3