Journal: Acta Neuropathologica Communications

Article Title: 5-HIAA induces neprilysin to ameliorate pathophysiology and symptoms in a mouse model for Alzheimer’s disease

doi: 10.1186/s40478-018-0640-z

Figure Lengend Snippet: 5-HIAA induces NEP expression via the MAP-kinase/ERK ½ pathway. a Down-regulatory effect of 5-HIAA on ERK ½, MEK ½ and GSK-3 expression in SH-SY5Y cells after 30 min treatment. The ERK pathway was down-regulated by 5-HIAA (mean decrease of ERK1–17% and ERK2–21%). GSK-3 α/β, a strategic factor for tau phosphorylation, was also significantly affected by the treatment (mean decrease of GSK-3α/β and GSK-3β − 15%), together with the transcription factor CREB (mean decrease − 40%). The statistical analysis of triplicate samples was done with a Student t-test. * p < 0.05, ** p < 0.005 by reference to non-treated cells (NT). b Effects of separated or combined treatments with the ERK ½ inhibitor SCH772984 and/or 5-HIAA on NEP expression in SH-SY5Y cells after 90 min treatment. NEP protein expression was stimulated by SCH772984 (26 +/− 4%) or 100 μM 5-HIAA (49 +/− 2%) tested separately. The combined treatment containing 100 μM 5-HIAA and SCH772984 showed a higher increase percentage of NEP expression (63 +/− 2%) compared to the separate treatments. The statistical analysis of triplicate samples was done by an ANOVA completed with a Bonferroni’s Multiple Comparison Test. ## p < 0.005, ### or *** p < 0.0001. * by reference to non-treated cells (NT), or # by reference to treatments with 5-HIAA. c Inhibitory action of 5-HIAA on AICD expression in SH-SY5Y cells. NH 4 Cl as positive control. The statistical analysis ( n = 5 in each group) was performed by an ANOVA completed with a Bonferroni’s Multiple Comparison Test. *** p < 0.0001 by reference to non-treated cells (NT). d Dose-response effect and EC 50 of SCH772984 and Trametinib on NEP expression in SH-SY5Y cells. Both MEK inhibitor (GSK 1120212, Trametinib) and ERK kinase inhibitor (SCH772984) induce NEP with an EC 50 of 14 +/− 4 and 27 +/− 3 nM, respectively. e Graphical summary of potential mechanisms involved in 5-HIAA-induced NEP expression. The MAP-kinase pathway is involved in NEP expression in SH-SY5Y cells: inhibition of MEK/ERK ½ phosphorylation increased NEP protein levels. Phosphorylation of GSK-3 is also down-regulated by 5-HIAA treatment. These effects may explain 5-HIAA-induced increase in Aβ peptide clearance

Article Snippet: ERK inhibitor SCH772984 (Carbosynth, Compton Berkshire, UK), MEK 1⁄2 inhibitor GSK 1120212 (Targetmol, USA) or GSK-3 inhibitor CHIR99021 (Sigma-Aldrich, France) was added 30 min before the measurement of NEP protein.

Techniques: Expressing, Phospho-proteomics, Comparison, Positive Control, Inhibition

Journal: Journal of Cell Communication and Signaling

Article Title: Microenvironment-sensing, nanocarrier-mediated delivery of combination chemotherapy for pancreatic cancer

doi: 10.1007/s12079-019-00514-w

Figure Lengend Snippet: a Flow cytometric analysis of cellular internalization of the nanoparticles labelled with FRET dyes by MIA PaCa-2 cells show a time dependent uptake (Black: control, Blue: dye labelled nanoparticles at 24 h, Red: at 48 h and Green at 72 h) (b) Detection of FRET associated fluorescence (540 nm) in the perinuclear space of MIA PaCa-2 cancer cells, when the cells were incubated with nanoparticles conjugated with FRET coupled dyes, i.e. AF 594 and AF 647. Arrow indicates the presence of nanoparticles around the nucleus which were stained with DAPI (c) Western blot data showing the cellular availability of SCH 772984, suppressing the phosphorylation of ERK (A: Encapsulated ERK inhibitor (1 nM), B: Free ERK inhibitor (1 nM), C: Bare Nanoparticles, D: Encapsulated ERK inhibitor (varying concentrations), E: Free ERK inhibitor (varying concentrations), F: Bare NCs (varying concentrations)

Article Snippet: Herein we report a pharmaceutical nanoformulation of GEM and an ERK inhibitor (SCH 772984) co-stabilized within a pH-sensing nanocarrier (NC, with a hydrodynamic diameter of 161 ± 5.0 nm).

Techniques: Control, Fluorescence, Incubation, Staining, Western Blot, Phospho-proteomics