Journal: Molecular Neurobiology

Article Title: Ptbp2 Alleviates Neuroinflammation and Blood–brain Barrier Disruption via Modulating Microglial Polarization in Ischemic Stroke

doi: 10.1007/s12035-026-05704-3

Figure Lengend Snippet: Ptbp2 modifies microglial polarization by downregulating the NF-κB pathway ( a- e ) Representative western blots of p-p65 and p65 and quantitative analysis of protein levels normalized to GAPDH ( n = 3). ( e-f ) Quantitative analysis of TNF-α levels normalized to GAPDH and representative western blots ( n = 3). ( g-l ) ELISA for evaluating TNF-α and IL-10 levels in the mouse blood, brain tissue, and BV-2 cells ( n = 6). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: TNF-α and IL-10 concentrations were determined using the quick ELISA kits (TNF-α, BOSTER EK0527; IL-10, BOSTER EK0417) following the supplier's instructions.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Thoracic Disease

Article Title: Mesenchymal stem cells transfected with anti-miRNA-204-3p inhibit acute rejection after heart transplantation by targeting C-X-C motif chemokine receptor 4 (CXCR4) in vitro

doi: 10.21037/jtd-21-1293

Figure Lengend Snippet: Expression of CXCR4 on MSC surface promotes macrophage polarization toward the M2 phenotype. (A) Relative expression of IL-10 and CD206 mRNA in macrophages co-cultured with MSCs transfected with CXCR4 shRNA and cDNA compared with that of their respective negative controls. (B) Arg-1 and CD206 protein abundance. (C) Concentration of IL-10 secreted by macrophages. All data are presented as mean ± SD. **P<0.01, ****P<0.0001, analyzed by Student’s t-test (n=3). CXCR4, C-X-C motif chemokine receptor 4; MSC, mesenchymal stem cell; IL-10, interleukin 10; mRNA, messenger RNA; shRNA, short hairpin RNA; cDNA, complementary DNA; SD, standard deviation.

Article Snippet: All the primers ( ) were designed and synthesized by GenePharma (Shanghai, China). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Primer sequence CXCR4 F: 5'- CTGAACCCCATCCTCTACGC-3' R: 5'- GAAGAGTGTCCACCCCGTTT-3' CD206 F: 5'-CCTATCTCTCCAACCACGGC-3' R: 5'-GGCAAGCAGGGAAGTACGAT-3' IL-10 F: 5'-TAACTGCACCCACTTCCCAG-3' R: 5'-TGGCAACCCAAGTAACCCTTAAA-3' miR-204-3p F: 5'-TGTTGCAGTGAGGGCAAGAA-3' R: 5'-GACCCTGGTTGCTTCAAGGA-3' GAPDH F: 5 ’ -ACAGCAACAGGGTGGTGGAC-3' R: 5'-TTTGAGGGTGCAGCGAACTT-3' U6 F: 5'-CAGCACATATACTAAAAAAGGAACG-3' R: 5'- ACGAATTTGCGTGTCATCC-3' Open in a separate window qRT-PCR, quantitative reverse transcription polymerase chain reaction; CXCR4, C-X-C motif chemokine receptor 4; CD206, Cluster of Differentiation 206; IL-10, interleukin-10; GAPDH, glyceraldehydes-3-phosphate dehydrogenase. caption a8 Primer sequences for qRT-PCR Western blot Cells were washed thrice with cold PBS and lysed on ice in radioimmunoprecipitation assay (RIPA) buffer supplemented with 1% Phosphatase Inhibitor Cocktail III (MedChem Express, Monmouth Junction, NJ, USA).

Techniques: Expressing, Cell Culture, Transfection, shRNA, Concentration Assay, Standard Deviation

Journal: Journal of Thoracic Disease

Article Title: Mesenchymal stem cells transfected with anti-miRNA-204-3p inhibit acute rejection after heart transplantation by targeting C-X-C motif chemokine receptor 4 (CXCR4) in vitro

doi: 10.21037/jtd-21-1293

Figure Lengend Snippet: MSCs transfected with anti-miR-204-3p promote macrophage polarization toward the M2 phenotype. (A) Relative expression of IL-10 and CD206 mRNA in macrophages co-cultured with MSCs. (B) Arg-1 and CD206 protein abundance. (C) Concentration of IL-10 secreted by macrophages. All data are presented as mean ± SD. *P<0.05, **P<0.01, ****P<0.0001, analyzed by Student’s t-test (n=3). MSCs, mesenchymal stem cells; mRNA, messenger RNA; IL-10, interleukin 10; SD, standard deviation.

Article Snippet: All the primers ( ) were designed and synthesized by GenePharma (Shanghai, China). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Primer sequence CXCR4 F: 5'- CTGAACCCCATCCTCTACGC-3' R: 5'- GAAGAGTGTCCACCCCGTTT-3' CD206 F: 5'-CCTATCTCTCCAACCACGGC-3' R: 5'-GGCAAGCAGGGAAGTACGAT-3' IL-10 F: 5'-TAACTGCACCCACTTCCCAG-3' R: 5'-TGGCAACCCAAGTAACCCTTAAA-3' miR-204-3p F: 5'-TGTTGCAGTGAGGGCAAGAA-3' R: 5'-GACCCTGGTTGCTTCAAGGA-3' GAPDH F: 5 ’ -ACAGCAACAGGGTGGTGGAC-3' R: 5'-TTTGAGGGTGCAGCGAACTT-3' U6 F: 5'-CAGCACATATACTAAAAAAGGAACG-3' R: 5'- ACGAATTTGCGTGTCATCC-3' Open in a separate window qRT-PCR, quantitative reverse transcription polymerase chain reaction; CXCR4, C-X-C motif chemokine receptor 4; CD206, Cluster of Differentiation 206; IL-10, interleukin-10; GAPDH, glyceraldehydes-3-phosphate dehydrogenase. caption a8 Primer sequences for qRT-PCR Western blot Cells were washed thrice with cold PBS and lysed on ice in radioimmunoprecipitation assay (RIPA) buffer supplemented with 1% Phosphatase Inhibitor Cocktail III (MedChem Express, Monmouth Junction, NJ, USA).

Techniques: Transfection, Expressing, Cell Culture, Concentration Assay, Standard Deviation