Journal: Stem Cell Research & Therapy
Article Title: Lumican from DESC spheroids enhances odontoblastic differentiation of SCAPs
doi: 10.1186/s13287-025-04849-7
Figure Lengend Snippet: Lumican enhances SCAPs’ odontoblastic differentiation via ITGβ1-ERK pathway ( a , b ) Western blot analysis and quantification of phosphorylation levels of JNK, p38 and ERK in SCAPs following Lumican treatment. Note the time-dependent increase in phosphorylation of both JNK and ERK. Control: 5% DMSO without inhibitor. c Fluorescence microscopy images revealed increased fluorescence intensity following Lumican treatment. d ALP and ARS staining showing that U0126 treatment most significantly abolished the pro-odontoblastic effects of Lumican on SCAPs. Control: 5% DMSO without inhibitor. e RT-qPCR demonstrating that Lumican-induced upregulation of odontogenesis-related markers Dspp , Dmp1 , Ocn and Nestin was most significantly reversed by U0126 treatment. Control: 5% DMSO without inhibitor. f , g Western blot analysis and quantification demonstrating that Lumican-induced upregulation of odontogenesis-related markers DSPP, DMP1 and OCN at both protein levels was reversed by U0126 treatment. Control: 5% DMSO without inhibitor ( n = 3; one-way ANOVA). h , i Western blot analysis and quantification showing increased ITGβ1 expression in Lumican-treated SCAPs. j Fluorescence microscopy images revealing co-localization (yellow) of ITGβ1 (red) and 6*His-tagged Lumican (green) in SCAPs membrane. k Co-IP assay confirming direct interaction between 6*His-tagged Lumican and ITGβ1. l , m Western blot analysis and quantification showing AIIB2 treatment did not affect ITGβ1 expression in SCAPs, but attenuated Lumican-enhanced ERK phosphorylation. n ALP and ARS staining showed that AIIB2 pretreatment counteracted the enhancing effect of Lumican on both early differentiation and late-stage mineralization. o RT-qPCR demonstrating that AIIB2 pretreatment counteracted the Lumican-induced upregulation of odontogenesis-related markers Dspp , Dmp1 , Ocn and Nestin . (p , q) Western blot analysis and quantification demonstrating that AIIB2 pretreatment counteracted the Lumican-induced upregulation of odontogenesis-related markers Dspp , Dmp1 , Ocn and Nestin
Article Snippet: After fixation, permeabilization, and blocking, the cells were incubated with anti-6*His antibody (66005-1-Ig, Proteintech, China, 1:200) and anti-integrin β1 (ITGβ1) antibody (12594-1-AP, Proteintech, China, 1:200) at 4 °C for 16 h. The cells were rinsed with PBS and treated with Alexa Fluor 488-conjugated (AF488, Beyotime, China, 1:200) and Alexa Fluor 647-conjugated secondary antibodies (A0473, Beyotime, China, 1:200) at room temperature for 1 h. The nuclei were counterstained with DAPI for 5 min.
Techniques: Western Blot, Phospho-proteomics, Control, Fluorescence, Microscopy, Staining, Quantitative RT-PCR, Expressing, Membrane, Co-Immunoprecipitation Assay