Journal: Molecular immunology

Article Title: Methamphetamine alters the TLR4 signaling pathway, NF-κB activation, and pro-inflammatory cytokine production in LPS-challenged NR-9460 microglia-like cells

doi: 10.1016/j.molimm.2020.03.013

Figure Lengend Snippet: (A) The expression of MyD88, IRAK1, TRAF6, IKK, NF-kB, p38, and IRF3 in microglia-like cells grown in absence (untreated) or presence of LPS, METH, MC, METH + LPS, and MC + LPS was determined by a Western blot analysis and a representative gel is shown. A density of 105 NR-9460 cells was treated with 25 μM METH or 1 μM MC for 2 h followed by an incubation with 10 μM LPS for 24 h. The housekeeping gene Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a control. The levels of expression of (B) MyD88, (C) IRAK1, (D) TRAF6, (E) IKK, (F) NF-kB, (G) p38, and (H) IRF3 were measured. Individual band intensities from the Western blot in panel A were quantified using ImageJ software. GAPDH was used to determine the relative intensity ratios shown in graphs B-G. Symbols (*, X, @, #, ψ, λ, Φ, &, %, and ε) indicate P value significance (P < 0.05) calculated using ANOVA and adjusted by use of the Tukey’s multiple comparison analysis. *, ψ, λ, Φ, and ε indicate significantly higher protein expression than in microglia from the untreated, LPS−, METH−, MC−, and METH + LPS-treated groups, respectively. X, @, #, %, and & indicate significantly lower protein expression than in microglia from the untreated, LPS−, METH−, MC− and METH + LPS-treated groups, respectively. Each gel was run 3X and similar results were obtained each time.

Article Snippet: Actin (dilution, 1:1,000; TF), a structural housekeeping protein and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; dilution, 1:1,000; BD), a cytoplasmic housekeeping protein, were used as a loading control to determine the relative intensity ratio.

Techniques: Expressing, Western Blot, Incubation, Software