Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: Actin (smooth muscle) , REAL650 , 50 , 130-123-363 , FITC , Miltenyi Biotec.

Techniques: Imaging

Journal: International Journal of Molecular Sciences

Article Title: Serpine1 Regulates the Enhanced Inhibitory Effect of CHIR99021 Combined with Fibroblast Growth Factor 2 on Myocardial Fibrosis After Myocardial Infarction in Mice

doi: 10.3390/ijms27041627

Figure Lengend Snippet: CHIR99021 combined with FGF2 inhibits collagen secretion and activation of cardiac fbroblasts. ( A ) The individual and combined effects of CHIR99021 and FGF2 led to significant changes in the morphology of cardiac fibroblasts. Scale bar: 100 μm. ( B ) Immunofluorescence staining of α-SMA, ColI, and ColIII in CFs treated with CHIR99021 and/or FGF2. Scale bar: 50 μm. ( C ) Quantitative analysis of α-SMA, ColI, and ColIII expression in CFs following treatment. n = 3. * p < 0.05, ** p < 0.01. ( D ) Western blotting analysis of α-SMA expression in CFs under different treatment conditions. n = 3. ** p < 0.01. ( E ) Quantification of hydroxyproline content in CFs treated with CHIR99021 and/or FGF2. n = 3. * p < 0.05.

Article Snippet: CFs were seeded into 96-well plates at 3000 cells/well and treated with CHIR99021 (10 μM), FGF2 (20 ng/mL), or both for 24 h. Immunofluorescence staining was performed for α-SMA, ColI, ColIII, Serpine1 and Cryab using the following antibodies: α-SMA (ET1607-53, HuaBio, 1:1000, Hangzhou, China), ColI (HA722517, HuaBio, 1:500, Hangzhou, China), and ColIII (HA720050, HuaBio, 1:500, Hangzhou, China), Serpine1 ( HA500124 , HuaBio, 1:100, Hangzhou, China) and Cryab (CY6897, Abways, 1:100, Shanghai, China).

Techniques: Activation Assay, Immunofluorescence, Staining, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Serpine1 Regulates the Enhanced Inhibitory Effect of CHIR99021 Combined with Fibroblast Growth Factor 2 on Myocardial Fibrosis After Myocardial Infarction in Mice

doi: 10.3390/ijms27041627

Figure Lengend Snippet: CHIR99021 combined with FGF2 inhibits the expression of genes related to fibrosis, n = 3. ( A ) Relative mRNA expression levels of fibrosis-related genes ( Acta2 , Col3a1 , Col1a1 and Serpine1 ) in CFs under different treatment conditions, as determined by qPCR. Gapdh was used as the internal control. * p < 0.05, ** p < 0.01. ( B ) Immunofluorescence staining showing the expression of α-SMA, ColIII, and Serpine1 in CFs. Scale bar: 100 μm. ( C ) Quantitative determination of hydroxyproline content in the culture supernatant of CFs following various treatments. * p < 0.05. ( D ) Western blotting analysis showing the protein expression levels of α-SMA, Col III, and Serpine1 in CFs treated with CHIR99021 and FGF2. ( E ) Quantitative analysis of the Western blot results. Scale bar: 100 μm. * p < 0.05, ** p < 0.01.

Article Snippet: CFs were seeded into 96-well plates at 3000 cells/well and treated with CHIR99021 (10 μM), FGF2 (20 ng/mL), or both for 24 h. Immunofluorescence staining was performed for α-SMA, ColI, ColIII, Serpine1 and Cryab using the following antibodies: α-SMA (ET1607-53, HuaBio, 1:1000, Hangzhou, China), ColI (HA722517, HuaBio, 1:500, Hangzhou, China), and ColIII (HA720050, HuaBio, 1:500, Hangzhou, China), Serpine1 ( HA500124 , HuaBio, 1:100, Hangzhou, China) and Cryab (CY6897, Abways, 1:100, Shanghai, China).

Techniques: Expressing, Control, Immunofluorescence, Staining, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Serpine1 Regulates the Enhanced Inhibitory Effect of CHIR99021 Combined with Fibroblast Growth Factor 2 on Myocardial Fibrosis After Myocardial Infarction in Mice

doi: 10.3390/ijms27041627

Figure Lengend Snippet: Role of Serpine1 in Cardiac Fibroblast Activation and Collagen Expression. ( A ) Serpine1 knockdown and overexpression on the morphological changes in cardiac fibroblasts. Scale bar: 100 μm. ( B ) Western blotting analysis of ColIII, α-SMA, and Col I protein expression in CFs following Serpine1 knockdown. n = 3. ** p < 0.01, *** p < 0.001. ( C ) Immunofluorescence analysis of Serpine1, α-SMA, ColIII and Cryab expression in CFs after Serpine1 knockdown and overexpression. Scale bar: 50 μm.

Article Snippet: CFs were seeded into 96-well plates at 3000 cells/well and treated with CHIR99021 (10 μM), FGF2 (20 ng/mL), or both for 24 h. Immunofluorescence staining was performed for α-SMA, ColI, ColIII, Serpine1 and Cryab using the following antibodies: α-SMA (ET1607-53, HuaBio, 1:1000, Hangzhou, China), ColI (HA722517, HuaBio, 1:500, Hangzhou, China), and ColIII (HA720050, HuaBio, 1:500, Hangzhou, China), Serpine1 ( HA500124 , HuaBio, 1:100, Hangzhou, China) and Cryab (CY6897, Abways, 1:100, Shanghai, China).

Techniques: Activation Assay, Expressing, Knockdown, Over Expression, Western Blot, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Serpine1 Regulates the Enhanced Inhibitory Effect of CHIR99021 Combined with Fibroblast Growth Factor 2 on Myocardial Fibrosis After Myocardial Infarction in Mice

doi: 10.3390/ijms27041627

Figure Lengend Snippet: CHIR99021 and FGF2 regulate Serpine1 through the TGF-β and FAK signaling pathway. ( A ) Western blot assay showing the changes in protein expression of fibroblast activation markers (ColIII; and α-SMA), Smad2/3 and FAK through the overexpression of Serpine1 in cardiac fibroblasts following the indicated treatments. ( B ) Statistical analysis of relative protein expression levels n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: CFs were seeded into 96-well plates at 3000 cells/well and treated with CHIR99021 (10 μM), FGF2 (20 ng/mL), or both for 24 h. Immunofluorescence staining was performed for α-SMA, ColI, ColIII, Serpine1 and Cryab using the following antibodies: α-SMA (ET1607-53, HuaBio, 1:1000, Hangzhou, China), ColI (HA722517, HuaBio, 1:500, Hangzhou, China), and ColIII (HA720050, HuaBio, 1:500, Hangzhou, China), Serpine1 ( HA500124 , HuaBio, 1:100, Hangzhou, China) and Cryab (CY6897, Abways, 1:100, Shanghai, China).

Techniques: Western Blot, Expressing, Activation Assay, Over Expression

Journal: Acta Pharmacologica Sinica

Article Title: A novel biphenyl compound IMB-S7 ameliorates hepatic fibrosis in BDL rats by suppressing Sp1-mediated integrin αv expression

doi: 10.1038/s41401-019-0325-6

Figure Lengend Snippet: a LX2 cells were treated with different concentrations of IMB-S7 for the indicated times, and cell viability was measured by the sulforhodamine B (SRB) assay. b , c LX2 cells were starved for 24 h and then treated with TGF-β1 (2 ng/mL) and IMB-S7 (0.2 mM or 0.5 mM) for an additional 24 h. Real-time PCR for TGFB1, COL1A1, and ACTA2 ( b ) and Western blot analysis for TGF-β1, COL1A1, α-SMA, p-Smad2/3, and Smad2 ( c ), * P < 0.05 vs the DMSO group; # P < 0.05 vs the TGF-β1 group. d , e The mRNA and protein as in b and c in liver samples of the BDL rat model were also detected by real-time PCR and Western blot ( n = 6 per group), * P < 0.05, ** P < 0.01 vs the sham group; # P < 0.05 vs the BDL group. Mouse primary HSCs were treated with IMB-S7 (0.2 or 0.5 mM) for 24 h. Real-time PCR for Tgfβ1 , Col1a1 , and Acta2 ( f ) and western blot analysis for TGF-β1, COL1A1, α-SMA, p-Smad2/3 and Smad2 ( g ). * P < 0.05 vs the control group.

Article Snippet: Anti-α-SMA and anti-COL1A1 antibodies were obtained from Abnova (Taipei, Chinese Taiwan).

Techniques: Sulforhodamine B Assay, Real-time Polymerase Chain Reaction, Western Blot, Control