Figures & data
Figure 1. An overall flowchart for the research methodology
Figure 2. After 100 ng/mL LPS treatment on microglia, protein expressions of inflammatory factors and microglial cell marker Iba1 were determined (n = 3). (a) Immunofluorescence detection of Iba1 expression. (b-d) ELISA detection of TNF-α, IL-6, and IL-1β expression. The scale bar is 50 μm. **, P < 0.01 compared with the control
Figure 3. miR-27b-3p promoted expressions of TNF-α, IL-6, and IL-1β in microglial cells treated with 100 ng/mL LPS (n = 3). (a) Expression of miR-27b-3p determined in transfected microglial cells. (b-d) TNF-α, IL-6, and IL-1β mRNA expression were determined by qPCR. (e) Western blot assay was used for detecting TNF-α, IL-6, and IL-1β protein expressions. The relative density was measured by Image J. mimic-NC: mimic negative control; si-NC: inhibitor negative control. *, p < 0.05; **, p < 0.01 compared to control
Figure 4. miR-27b-3p accelerated apoptosis and Iba1 expression inmicroglial cells treated with 100 ng/mL LPS (n = 3). (a) Flow cytometry for detecting apoptosis of miR-27b-3p in microglia. (b) Immunofluorescence for detecting Iba1 expression. **, P < 0.01 compared with the NC control
Figure 5. A20 target gene of miR-27b-3p (n = 3). (a) Overexpressed miR-129-5p downregulated the luciferase activity of only wild-type A20 rather than mutant A20 in 293 T cells. (b) qPCR detection on A20 following miR-27b-3p overexpression in microglial cells. (c) Western blot assay was employed to determine A20 expression following overexpressed miR-27b-3p in microglial cells. (d) Immunofluorescence detection of A20 in microglial cells by overexpressed miR-27b-3p. The scale bar is 20 μm. ##, p < 0.01 compared with the Control mimics
Figure 6. A20 inhibited TNF-α, IL-6, and IL-1β expressions in microglial cells treated with 100 ng/mL LPS (n = 3). Both qPCR assay (a) and western blot assay (b) were adopted for the detection of A20 mRNA and protein expression, respectively. (c-e) mRNA levels of TNF-α, IL-6, and IL-β were subjected to qPCR. (f) Protein levels of TNF-α, IL-6, and IL-β were subjected to western blot. The relative density of TNF-α, IL-6, IL-1β, and A20 proteins was measured by Image J. (g) Apoptosis of microglia was detected by flow cytometry. OE-NC: overexpression negative control; si-NC: siRNA negative control. **, p < 0.01 compared with the negative control
