Journal: PLOS One

Article Title: Single-cell profiling unveils key regulators of skeletal stem cells in chicken and human embryonic limb development

doi: 10.1371/journal.pone.0346514

Figure Lengend Snippet: Expression of COL5A2, COL1A2, PRRX1, and TGFβ2 was detected in the periosteal region and the primary ossification centers. A. Merged and single-channel images of COL5A2 (red) and DAPI (blue) staining. B. Merged and single-channel images of COL1A2 (red) and DAPI (blue) staining. C. Merged and single-channel images of PRRX1 (red) and DAPI (blue) staining. D. Merged and single-channel images of TGFβ2 (red) and DAPI (blue) staining. All experiments were independently repeated at least three times. Scale bars in snapshot image, 200 μm; scale bars in magnified images, 50 μm.

Article Snippet: The following antibodies were used: anti-COL5A2 antibody ( GB111012 , 1:500, Servicebio, Wuhan, China), anti-COL1A2 antibody (AF7001, 1:200, Affinity Biosciences, Jiangsu, China), anti-PRRX1 antibody (DF4274, 1:200, Affinity Biosciences, Jiangsu, China), anti-TGFβ2 antibody (AF0260, 1:100, Affinity Biosciences, Jiangsu, China), and Cy3-conjugated goat anti-rabbit IgG (GB21303, 1:300, Servicebio, Wuhan, China).

Techniques: Expressing, Staining

Journal: Inflammation

Article Title: A Novel Insight into Chronic Pancreatitis Pathogenesis: the USP1/ITGB5 Axis-Mediated Stellate Cell Activation

doi: 10.1007/s10753-025-02434-x

Figure Lengend Snippet: Proteomics reveals impairment of ECM degradation in PSCs with USP1 knockdown. A , B . USP1 expression was estimated in PSCs treated with different concentration of TGF-β1 (n = 3). C , D . The USP1 expression after lentiviral transfection was detected by real-time PCR ( C ) and western blot ( D ) ( n = 3). E Heap map of differentially expressed proteins (DEPs) in label-free proteomic ( n = 4). F . GO enrichment analysis of DEPs associated with collagen fibers. G . Heat map of collagen protein (n = 4). H . The protein expression of COL1A1, COL1A2, and FN in the PSCs was determined by western blot ( n = 3). I . Immunofluorescence staining of COL1A1 in the PSCs ( n = 3). Bar: 50 μm. J . Immunofluorescence staining of α-SMA. Bar: 50 μm. K . Quantitative analysis of the fluorescence intensity of COL1A1 ( I ) and α-SMA (J) ( n = 3). **, p < 0.01; ***, p < 0.001

Article Snippet: COL1A2 ITGB5 PI3K p-PI3K AKT , 343,277 SC398214 AF6242 AF3242 AF6261 , 1: 500 1:1000 1:1000 1:1000 1:1000 , Zenbio Santa Cruz Affinity Affinity Affinity , WB WB/IP WB WB WB , .

Techniques: Knockdown, Expressing, Concentration Assay, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Staining, Fluorescence

Journal: Inflammation

Article Title: A Novel Insight into Chronic Pancreatitis Pathogenesis: the USP1/ITGB5 Axis-Mediated Stellate Cell Activation

doi: 10.1007/s10753-025-02434-x

Figure Lengend Snippet: USP1 knockdown promotes ITGB5 ubiquitination-mediated degradation. ( A ) Venn plot showing a total of 2015 proteins bind to USP1 under TGF-β1 stimulation. (B) KEGG pathway analysis and GO enrichment analysis pair of 2015 proteins in ( A ). ( C ) Label-free proteomics combined with IP-LC/MS to analysis the USP1 downstream target. The intersection of differential downregulated expressed protein of label-free proteomics, IP-LC/MS, and the upregulated genes of GSE41418 dataset. ( D ) The ITGB5-associated PPI was established via the String database. E-F. The expression of ITGB5 of the PSCs (E) ( n = 3) and pancreatic tissues ( F ) ( n = 6) was estimated by western blot. G . Co-IP shows the interaction of USP1 and ITGB5 in the PSCs ( n = 3). H . Ubiquitination detection of ITGB5 in the PSCs were measured by Co-IP ( n = 3). **, p < 0.01; ***, p < 0.001.

Article Snippet: COL1A2 ITGB5 PI3K p-PI3K AKT , 343,277 SC398214 AF6242 AF3242 AF6261 , 1: 500 1:1000 1:1000 1:1000 1:1000 , Zenbio Santa Cruz Affinity Affinity Affinity , WB WB/IP WB WB WB , .

Techniques: Knockdown, Ubiquitin Proteomics, Liquid Chromatography with Mass Spectroscopy, Expressing, Western Blot, Co-Immunoprecipitation Assay

Journal: Inflammation

Article Title: A Novel Insight into Chronic Pancreatitis Pathogenesis: the USP1/ITGB5 Axis-Mediated Stellate Cell Activation

doi: 10.1007/s10753-025-02434-x

Figure Lengend Snippet: ITGB5 knockdown inhibits PSC activation by inhibiting the PI3K/AKT pathway. A . The ITGB5 expression in the PSCs after lentiviral transfection was detected by western blot ( n = 3). B , C . The expression of COL1A1 ( B ) and FN ( C ) was detected by western blot ( n = 3). D . IF staining showed the α-SMA expression ( n = 3). Bar: 50 μm. E . KEGG analysis of DEPs of label-free proteomics. F , G . The phosphorylation levels of PI3K and AKT and the PI3K and AKT protein expression was measured by western blot (n = 3). H , I . The expression of COL1A1 and FN was detected by western blot ( n = 3). J . α-SMA expression in the PSCs was determined by IF staining (n = 3). Bar: 50 μm. **, p < 0.01; ***, p < 0.001.

Article Snippet: COL1A2 ITGB5 PI3K p-PI3K AKT , 343,277 SC398214 AF6242 AF3242 AF6261 , 1: 500 1:1000 1:1000 1:1000 1:1000 , Zenbio Santa Cruz Affinity Affinity Affinity , WB WB/IP WB WB WB , .

Techniques: Knockdown, Activation Assay, Expressing, Transfection, Western Blot, Staining, Phospho-proteomics

Journal: Inflammation

Article Title: A Novel Insight into Chronic Pancreatitis Pathogenesis: the USP1/ITGB5 Axis-Mediated Stellate Cell Activation

doi: 10.1007/s10753-025-02434-x

Figure Lengend Snippet: ITGB5 overexpression increased extracellular matrix. ( A ) ITGB5 expression in the PSCs was detected by western blot ( n = 3). ( B ) COL1A1 expression in the PSCs was detected by western blot ( n = 3). (C) FN expression in the PSCs was detected by western blot ( n = 3). ( D ) Immunofluorescence staining of α-SMA in the PSCs ( n = 3). Bar: 50 μm. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: COL1A2 ITGB5 PI3K p-PI3K AKT , 343,277 SC398214 AF6242 AF3242 AF6261 , 1: 500 1:1000 1:1000 1:1000 1:1000 , Zenbio Santa Cruz Affinity Affinity Affinity , WB WB/IP WB WB WB , .

Techniques: Over Expression, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Inflammation

Article Title: A Novel Insight into Chronic Pancreatitis Pathogenesis: the USP1/ITGB5 Axis-Mediated Stellate Cell Activation

doi: 10.1007/s10753-025-02434-x

Figure Lengend Snippet: Graphical summary of the possible mechanisms of USP1/ITGB5 axis in CP progression. USP1 deubiquitinates and stabilizes ITGB5, upregulates ITGB5 expression, and ITGB5 activated the PI3K/AKT pathway and subsequently activates PSCs, leading to pancreatic fibrotic injury in CP. Arrow: promotion; “tee” head: inhibition

Article Snippet: COL1A2 ITGB5 PI3K p-PI3K AKT , 343,277 SC398214 AF6242 AF3242 AF6261 , 1: 500 1:1000 1:1000 1:1000 1:1000 , Zenbio Santa Cruz Affinity Affinity Affinity , WB WB/IP WB WB WB , .

Techniques: Expressing, Inhibition