Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 5. Expression changes of key GBM proteins and their response to FN1 Variants in TBMN. Immunofluorescence experiments assessed the expression of Laminin α5β2, COL4A3/4/5 and Integrin β1 in the GBM. The findings revealed non-homogeneous linear structure expression of these key GBM proteins in the presence of FN1 variants, indicating their potential involvement in the pathogenesis of TBMN. HC = healthy control.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Expressing, Immunofluorescence, Control

Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 6. Abnormal co-localization of FN1 variants with Integrin β1 and its impact on other GBM components. Immunofluorescence studies investigated the rela tionship between FN1 variants and the expression patterns of GBM components. The findings reveal abnormal co-localization of FN1 variants with Integrin β1, accompanied by reduced co-localization of other GBM components. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Immunofluorescence, Expressing

Journal: International journal of biological macromolecules

Article Title: Aberrant serum-derived FN1 variants bind to integrin β1 on glomerular endothelial cells contributing to thin basement membrane nephropathy.

doi: 10.1016/j.ijbiomac.2024.136282

Figure Lengend Snippet: Fig. 7. Competitive Binding of FN1 Variants to Integrin β1. (A and B) Duolink proximity ligation assay and Co-IP were employed to analyze the protein interaction between FN1 variants and Integrin β1. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Incubating primary antibodies overnight at 4 ◦C, with specific primary antibodies against HA (1:2000, ab9110, Abcam), Integrin β1 (1:1000, 12,594–1-AP, Proteintech), β-tubulin (1:3000, ab0039, Abways), Laminin α5 (1:1000, ab210957, Abcam), Laminin β2 (1:1000, ab210956, Abcam), Laminin γ1 (1:1000, ab233389, Abcam) in Tris-Buffered Saline Tween-20 (TBST) containing 5 % skim milk.

Techniques: Binding Assay, Proximity Ligation Assay, Co-Immunoprecipitation Assay

Journal: Cancer Medicine

Article Title: The expression and role of SUZ12 in lung adenocarcinoma

doi: 10.1002/cam4.70190

Figure Lengend Snippet: The effect of SUZ12 on metastasis related genes expression was tested by qRT‐PCR and western blotting. sh‐SUZ12 increased MMP1/2/3/9 mRNA expression (A) and TIMP1/2 protein expression (B), while decreased MMP14 mRNA expression (A), MMP1/9/14 (B), TIMP3 (B), and ITGB1/5(F) protein expression, without significantly effected ITGBs mRNA expression (E). oe‐SUZ12 increased MMP14 mRNA (C) and protein (D) expression, while decreased TIMP2 mRNA expression (C) and TIMP1/2 (D) protein expression.

Article Snippet: The list of primary antibodies: SUZ12 (1 μg/mL, Abcam Cambridge, cat no: ab12073), CDK2 (1:1000; Proteintech, USA, cat. no. 10122‐1‐AP), CDK3 (1:2000; Proteintech, USA, cat. no. 55103‐1‐AP), CDK6 (1:2000; Proteintech, USA, cat. no. 14052‐1‐AP), cyclin D1 (1:5000; Proteintech, USA, cat. no. 26939‐1‐AP), cyclin E1 (1:1000; Proteintech, USA, cat. no. 11554‐1‐AP), p18 (1:1000, BOSTER China, cat. no. M03299‐1), p19 (1:1000, BOSTER China, cat. no. MA1075), p53 (1:5000; Proteintech, USA, cat. no. 60283‐2‐Ig), p‐p53 (1:2000; Proteintech, USA, cat. no. 28961‐1‐AP), p57 (1:1000, BOSTER China, cat. no. BM4129), Rb (1:1000, BOSTER China, cat. no. BM4500), pRb (1:1000, BOSTER China, cat. no. BM4338), Bcl‐2 (1:1000; Proteintech, USA, cat. no. 26593‐1‐AP), Bax (1:2000; Proteintech, USA, cat. no. 50599‐2‐lg), E‐cadherin (1:5000; Proteintech, USA, cat. no. 20874‐1‐AP), N‐cadherin (1:3000; Proteintech, USA, cat. no. 22018‐1‐AP), vimentin (1:4000; Proteintech, USA, cat. no. 10366‐1‐AP), MMP1 (1:1000, BOSTER China, cat. no. A00733‐1), MMP2 (1:500, BOSTER China, cat. no. BM4075), MMP9 (1:1000, BOSTER China, cat. no. PB0709), MMP14 (1:1000, BOSTER China, cat. no. BM4119), TIMP1 (1:1000, Bioss China, cat. no. bs‐0415R), TIMP2 (1:1000, Bioss China, cat. no. bs‐10395R), TIMP3 (1:1000; Proteintech, USA, cat. no. 10858‐1‐AP), ITGB1 (1:1000, BOSTER China, cat. no. BM4308), ITGB3 (1:1000, BOSTER China, cat. no. BA1670), ITGB5 (1:1000, BOSTER China, cat. no. A04201‐1), nm23 (1:1000, BOSTER China, cat. no. BA3787), PD‐L1 (1:3000; Proteintech, USA, cat. no. 66248‐1‐Ig), and β‐actin (1:5000; Proteintech, USA, cat. no. 66009‐1‐Ig).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Neural Regeneration Research

Article Title: Pre-clinical study of human umbilical cord mesenchymal stem cell transplantation for the treatment of traumatic brain injury: safety evaluation from immunogenic and oncogenic perspectives

doi: 10.4103/1673-5374.317985

Figure Lengend Snippet: Identification, immunogenicity and immunomodulation of huMSCs . (A) The huMSCs cultured in our experiment. The huMSCs were spindle-shaped and grew in a whorl pattern. Original magnification 100×, scale bars: 200 μm. (B) Flow cytometry detection of CD29, CD44, CD73 and CD105. The rate of positive expression for all factors was greater than 95%. (C) Detection of huMSC HLAII expression in PBMCs and huMSCs by western blot. No HLAII expression band was detected in the huMSCs. (D) Relative expression of HLAII in PBMCs and huMSCs. The bar graph shows no HLAII expression in huMSCs. (E) PCR amplification curves of HLA-DPA1 , HLA-DQA1 and HLA-DRA1 genes in PBMCs and huMSCs. (F) Relative expression of HLA-DPA1 , HLA-DQA1 and HLA-DRA1 genes in PBMCs and huMSCs. PCR results showed no expression of HLA-DPA1 , HLA-DQA1 or HLA-DRA1 genes in huMSCs. (G) Expression curves of serum IL-6, IL-10, IL-12, TGF-β, and TNF-α detected by ELISA method ( n = 5 rats per group). Compared with the TBI group, the serum pro-inflammatory cytokines IL-6, IL-12 and TNF-α in the Tail Vein and In Situ group were lower (### P < 0.001), whereas the inflammation inhibiting factors IL-10 and TGF-β were much higher (### P < 0.001), indicating that huMSCs exert good immunoregulatory effects. Data are expressed as the mean ± SD. *** P < 0.001, vs . PBMCs (one-way analysis of variance followed by least significant difference test). ELISA: Enzyme-linked immunosorbent assay; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; HLAII: human leukocyte antigen II; huMSCs: human umbilical cord mesenchymal stem cells; IL: interleukin; PBMCs: peripheral blood mononuclear cells; PCR: polymerase chain reaction; TBI: traumatic brain injury; TGF-β: transforming growth factor beta; TNF-α: tumor necrosis factor alpha.

Article Snippet: The sections were thereafter incubated overnight at 4°C with primary antibodies, including rabbit anti-human CD29 polyclonal antibody (Cat# PB9086, 1:100, Boster), proliferating cell nuclear antigen (PCNA) mouse monoclonal antibody (Cat# 2586S, 1:4000, CST, Danvers, MA, USA), Caspase 3 (CASP3) rabbit polyclonal antibody (Cat# 19677-1-AP, 1:200, Proteintech) and F4/80 rabbit polyclonal antibody (Cat# 28463-1-AP, 1:200, Proteintech), diluted in 1% bovine serum albumin containing PBS.

Techniques: Immunopeptidomics, Cell Culture, Flow Cytometry, Expressing, Western Blot, Amplification, Enzyme-linked Immunosorbent Assay, In Situ, Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Pre-clinical study of human umbilical cord mesenchymal stem cell transplantation for the treatment of traumatic brain injury: safety evaluation from immunogenic and oncogenic perspectives

doi: 10.4103/1673-5374.317985

Figure Lengend Snippet: Sites of accumulation of huMSCs in vivo in TBI rats . (A) Live imaging of the primary organs of experimental rats. (B, C) The average fluorescence intensity in the brain, liver and lung in the In Situ (B) and Tail Vein (C) groups. Data are expressed as the mean ± SD ( n = 5 rats at each time point). # P < 0.05, ## P < 0.01, vs . TBI group (one-way analysis of variance followed by least significant difference test). (D, E) CD29 (red, stained by CoraLite594) immunoreactivity in CFSE-labeled huMSCs in brain tissue (immunofluorescence staining, original magnification 200×, scale bars: 100 μm). CD29 and CFSE co-labeled huMSCs were identified in the In Situ group on days 1, 3 and 7 (D) in the brain lesions. A few CFSE-labeled huMSCs in the Tail Vein group were also observed in the brain lesions on days 1 and 3 (E). CFSE: Carboxyfluorescein succinimidyl ester; CON: TBI group; DAPI: 4′,6-diamidine-2′-phenylindole dihydrochloride; huMSCs: human umbilical cord mesenchymal stem cells; TBI: traumatic brain injury.

Article Snippet: The sections were thereafter incubated overnight at 4°C with primary antibodies, including rabbit anti-human CD29 polyclonal antibody (Cat# PB9086, 1:100, Boster), proliferating cell nuclear antigen (PCNA) mouse monoclonal antibody (Cat# 2586S, 1:4000, CST, Danvers, MA, USA), Caspase 3 (CASP3) rabbit polyclonal antibody (Cat# 19677-1-AP, 1:200, Proteintech) and F4/80 rabbit polyclonal antibody (Cat# 28463-1-AP, 1:200, Proteintech), diluted in 1% bovine serum albumin containing PBS.

Techniques: In Vivo, Imaging, Fluorescence, In Situ, Staining, Labeling, Immunofluorescence