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: The inhibitors and antibodies used in this study included: U0126 (HY-12031 A, MCE, China, 10 μM), SP600125 (HY-12041, MCE, China, 10 μM), AIIB2 (Ab03450-6.1, Absolute, China, 1ug/ml), RGD (HY-P0278, MCE, China, dissolve in 5% DMSO, 5 μM), DMP1 (SC-73633, Santa Cruz, USA, 1:200), DSPP (SC-73632, Santa Cruz, USA, 1:200), OCN (SC-365797, Santa Cruz, USA, 1:100), ITGβ1 (12594-1-AP, Proteintech, China, 1:500), ERK (#4695, CST, USA, 1:1000), p-ERK (#4370, CST, USA, 1:2000), JNK (ab179461, Abcam, UK,1:1000), p-JNK (ab124956, Abcam, UK, 1:5000), p-38 ( AWA48968 , Abiowell, China, 1:1000), p-p38 ( AWA48964 , Abiowell, China, 1:1000), GAPDH (10494-1-AP, Proteintech, China, 1:5000), ACTIN (66009-1-Ig, Proteintech, China, 1:5000), HRP-conjugated goat anti-Rabbit Secondary Antibody (AWS0002, Abiowell, China, 1:5000), HRP-conjugated goat anti-Mouse Secondary Antibody (AWS0001, Abiowell, China, 1:5000).

Techniques: Western Blot, Phospho-proteomics, Control, Fluorescence, Microscopy, Staining, Quantitative RT-PCR, Expressing, Membrane, Co-Immunoprecipitation Assay