Journal: PeerJ

Article Title: Melatonin increases growth properties in human dermal papilla spheroids by activating AKT/GSK3β/β-Catenin signaling pathway

doi: 10.7717/peerj.13461

Figure Lengend Snippet: (A–E) HDP cells were treated with 0.75 and 1 mM melatonin for 48 h. (A–D) mRNA levels of ALP, VCAN, BMP2, and WNT5A were assessed by qRT-PCR and normalized against GAPDH . (E) Protein levels of these signature genes were assessed by western blotting and β-actin served as a loading control. (F–H) HDP cells were cotreated with 1 mM melatonin with or without 50 µM luzindole for 48 h. (F and G) mRNA levels of WNT5A and BMP2 were assessed by qRT-PCR and GAPDH served as an endogenous control. (H) Protein levels of WNT5A and BMP2 were assessed by western blotting and β-actin served as a loading control. Data are presented as the mean ± SD of three independent experiments. (A–D) One-way ANOVA, followed by Turkey’s post hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001 versus DMSO treated control (A, p = 0.0401; B, p = 0.0025; C, p = 0.012, and p = 0.003 respectively), and versus melatonin alone treatment control (F, p = 0.0008, p = 0.0005, and p = 0.0038; G, p = 0.0428, and p = 0.0058 respectively).

Article Snippet: Antibodies against protein kinase B (AKT) (#9272), phosphorylated AKT at serine 473 residue (#9271), phosphorylated AKT at threonine 308 residue (#9275), protein kinase A (PKA) (#4782), phosphorylated PKA at threonine 198 residue (#4781), extracellular signal-regulated kinase (ERK) (#9102), phosphorylated ERK at threonine 202/tyrosine 204 residues (#9101), p38 (#9212), phosphorylated p38 at threonine 180/tyrosine 182 residues (#9211), SRC (#2108), phosphorylated SRC at tyrosine 416 residue (#2101), glycogen synthase kinase 3β (GSK-3β) (#9315), phosphorylated GSK-3β at serine 9 residue (#9323), β-catenin (#9562S), phosphorylated β-catenin at serine 33/37/threonine 41 residues (#9561S), and WNT5a (#2392) were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Control