Fig. 5

Higher levels of proinflammatory cytokines in progranulin silenced RAW264.7 cells. a Cell viability of RAW264.7 cells after transfection in siControl or siGrn mice and exposure to hypoxia. Data are the means ± SEMs (n = 6). ^*P <0.05 vs. siControl; ^##P <0.01 vs. siControl + Hypoxia (Welch’s t-test). b Representative western blots showing immunoreactivity against TNF-α, C3, IL-1β, CCL2, and β-actin. c Quantitative analysis of expression level of TNF-α, C3, IL-1β, and CCL2 in RAW264.7 cells. Data are the means ± SEMs (n = 4). ^*P <0.05; ^**P <0.01 vs. siControl; ^#P <0.05, ^##P <0.01 vs. siControl + Hypoxia (TNF-α and C3: Dunn-Bonferroni post hoc method following a significant Kruskal-Wallis tests, IL-1β and CCL2: one-way ANOVA followed by Tukey’s tests). d Immunofluorescence staining of RAW264.7 cells with anti-iNOS (green) and anti-CD68 (red) antibodies. Nuclei were stained with Hoechst 33342 (blue). Scale bar: 10 μm