Journal: Mediators of Inflammation

Article Title: Amphiregulin Regulates Phagocytosis-Induced Cell Death in Monocytes via EGFR and Matrix Metalloproteinases

doi: 10.1155/2018/4310419

Figure Lengend Snippet: Neonatal monocytes display elevated pro-AREG surface expression and increased release of AREG in response to E. coli infection. PBMO and CBMO were incubated with E. coli for 1 h, extracellular bacteria were removed, and cells were cultivated for 24 h in total. (a) Immunocytological staining shows AREG protein expression (green) in monocytes. Texas Red-X phalloidin was applied to visualize F-actin in the cytoskeleton (red). (b) Pro-AREG surface expression in uninfected and E. coli -infected monocytes was quantified by using flow cytometry ( n = 5). Representative dot plots show gating strategy and cutoff value for AREG expression. (c) sAREG levels in the supernatant of monocytes were quantified by using ELISA ( n = 5). Data are shown as means + SD. Statistical analysis was performed using one-way ANOVA with Bonferroni's multiple comparisons test (ns: not significant, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.001).

Article Snippet: For detection of FasL, Human FasL PicoKine ELISA Kit (Boster Biological Technology, Pleasanton, USA) was used according to the manufacturer's instructions.

Techniques: Expressing, Infection, Incubation, Bacteria, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Amphiregulin Regulates Phagocytosis-Induced Cell Death in Monocytes via EGFR and Matrix Metalloproteinases

doi: 10.1155/2018/4310419

Figure Lengend Snippet: AREG mediates increased shedding of memFasL through engagement with EGFR. PBMO and CBMO were infected as described in . AREG stimulation and EGFR inhibitor treatment was started 1 h prior to infection and was maintained during cultivation. (a) memFasL surface expression in uninfected and E. coli -infected monocytes was analyzed in response to AREG stimulation and neutralization of EGFR by using flow cytometry ( n = 5). AREG stimulation significantly decreased FasL levels in infected PBMO but not CBMO. Neutralization of EGFR resulted in a complete rescue of infection-induced memFasL presentation. Representative dot plots show gating strategy and cutoff value. (b) sFasL levels in the supernatant of uninfected and E. coli -infected monocytes were quantified in response to AREG stimulation and neutralization of EGFR by using ELISA ( n = 3). AREG significantly increased sFasL levels in infected PBMO but not CBMO, while neutralization of EGFR abolished the effect. Data are shown as means + SD. Statistical significance was analyzed using two-way ANOVA with Bonferroni's multiple comparisons test ( ∗∗ p < 0.01, ∗∗∗ p < 0.005, and ∗∗∗∗ p < 0.001).

Article Snippet: For detection of FasL, Human FasL PicoKine ELISA Kit (Boster Biological Technology, Pleasanton, USA) was used according to the manufacturer's instructions.

Techniques: Infection, Expressing, Neutralization, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Amphiregulin Regulates Phagocytosis-Induced Cell Death in Monocytes via EGFR and Matrix Metalloproteinases

doi: 10.1155/2018/4310419

Figure Lengend Snippet: EGFR-dependent gelatinase activation mediates increased shedding of memFasL. PBMO and CBMO were infected as described in . Stimulation with AREG and gelatinase inhibition by CHX treatment was started 1 h prior to infection and was maintained during cultivation. (a) memFasL surface expression in uninfected and E. coli -infected monocytes was analyzed in response to AREG stimulation and gelatinase inhibition by using flow cytometry ( n = 4). Gelatinase inhibition prevented the decrease in memFasL presentation on monocytes triggered by AREG. (b) sFasL levels in the supernatant of uninfected and E. coli -infected monocytes were quantified in response to AREG stimulation and CHX treatment by using ELISA ( n = 3). Gelatinase inhibition was found to prevent the increase in sFasL levels triggered by AREG. Data are shown as means + SD. Statistical significance was analyzed using two-way ANOVA with Bonferroni's multiple comparisons test ( ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.001).

Article Snippet: For detection of FasL, Human FasL PicoKine ELISA Kit (Boster Biological Technology, Pleasanton, USA) was used according to the manufacturer's instructions.

Techniques: Activation Assay, Infection, Inhibition, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay