Journal: bioRxiv

Article Title: Nascent extracellular matrix converts biomaterial cues into cell fate decisions

doi: 10.64898/2026.01.10.698826

Figure Lengend Snippet: a. Timeline and schematic showing the blocking of cell adhesion to nECM using an integrin β1 (ITGB1) function perturbing antibody for 1 hour pre-embedding and for 7 consecutive days . b. Representative fluorescent images of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in high and low modification hydrogels at day 7 (scale bar = 100 μm). c. Quantification of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in high and low modification hydrogels at day 7 (low-Ctrl: n = 23 ROIs, N = 3; low-ITGB1i: n = 30 ROIs, N = 3; high-Ctrl: n = 23 ROIs, N = 3; low-ITGB1i: n = 27 ROIs, N = 3). d. Representative fluorescent images of Sox9 immunofluorescence (dashed line outlines nuclei) of chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in low and high modification hydrogels at day 7 (scale bar = 10 μm). e. Quantification of Sox9 nucleus-to-cytoplasm (NC) ratio of chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in low and high modification hydrogels at day 7 (low-Ctrl: n = 138 cells, N = 4; low-ITGB1i: n = 133 cells, N = 4; high-Ctrl: n = 124 cells, N = 4; high-ITGBi: n = 138 cells, N = 4). a-e. N = number of independent experiments ****p < 0.0001, *p < 0.05, error bar = standard deviation, ns: no significant difference by two-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: For perturbation studies, media was supplemented with either an anti-CD44 antibody (DSHB, H4C4, 2.5 μg/mL, day 0-3) or anti-integrin β1 antibody (anti-ITGB1, DSHB, AIIB2, 2.5 μg/mL, day 0-7).

Techniques: Blocking Assay, Cell Culture, Inhibition, Modification, Immunofluorescence, Standard Deviation

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

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