Journal: Biology direct

Article Title: RGS5 maintaining vascular homeostasis is altered by the tumor microenvironment.

doi: 10.1186/s13062-023-00437-y

Figure Lengend Snippet: Fig. 5 High RGS5 expression is associated with increased tumor vasculature in the patients with breast cancer. (A) Representative immunohistochemical staining for RGS5, α-SMA and CD34 expression in TNBC and adjacent normal tissues. Scale bar = 100 μm. Right: Quantification of the immunohistochemistry (n = 50). (B) Western blot and quantification of α-SMA and RGS5 in TNBC tissues. (C) Representative immunofluorescence images of RGS5 and α-SMA in TNBC tissues. Scale bar = 100 μm. The data were presented as the mean ± SD

Article Snippet: After blocking with 5% milk in TBST, the membranes were incubated with primary antibodies against VCAM-1 (1:1000, Epitomics), RGS5 (1:500, Proteintech), MCP-1(1:200, Santa Cruz), C3 (1:250, Abcam), CD74 (1:500, Abcam), Lyz2 (1:200, Proteintech) α-SMA(1:1000, Proteintech), Caspase-1 (1:200, Proteintech), GSDMD (1:200, Abcam) or β-actin (1:1000, Proteintech) at 4°C overnight, and then with the HRPconjugated secondary antibody for 1 h. The blots were visualized using Image Quant LAS 4000 detection system (GE Healthcare).

Techniques: Expressing, Immunohistochemical staining, Staining, Immunohistochemistry, Western Blot, Immunofluorescence

Journal: iScience

Article Title: The role of long noncoding RNA Nron in atherosclerosis development and plaque stability

doi: 10.1016/j.isci.2022.103978

Figure Lengend Snippet:

Article Snippet: Primary antibodies specific for F4/80 (ab100790, Abcam), α-SMA (ab240654, Abcam) or VEGFA (ab1316, Abcam) was used.

Techniques: Virus, Recombinant, Plasmid Preparation, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Silver Staining, In Vitro, TUNEL Assay, Software

Journal: Scientific Reports

Article Title: The antifibrotic effect of Vildagliptin and Diaminodiphenyl Sulfone in murine schistosomiasis mansoni

doi: 10.1038/s41598-025-91955-4

Figure Lengend Snippet: Effect of Vilda, DDS, and Vilda/DDS on α-SMA and MMP-9 as fibrotic markers on S. mansoni- infected mice. ( a ) Photomicrographs of immunohistochemical staining (IHC) of hepatic tissues with alpha-smooth muscle actin (α-SMA) (× 200, scale bar = 100µm) as follows: liver sections of normal control and normal treated groups (Control, Vilda, DDS, and Vilda/DDS) marked scarce hepatocytes with positive cytoplasmic reactivity to α-SMA antibody (arrow). Liver sections from (Infected to Inf + PZQ) highlighted positive expression of α-SMA along ova of Schistosoma (arrowhead), fibroblast encircling granuloma (circle), inflammatory cells with nuclear expression (cube), and surrounding hepatocytes with cytoplasmic reactivity (arrow). The expressions were graded as intense in the Infected group, high in Inf + DDS group , moderately high in Inf + PZQ group , moderately low in Inf + Vilda/DDS group , and few in Inf + Vilda group. ( b ) Quantitative analysis of hepatic α-SMA IHC expression; (c) Hepatic level of matrix metallopeptidase-9 (MMP-9) as a fibrotic marker . Data are presented as mean ± SEM, (n = 6) and analyzed by one-way ANOVA followed by Tukey multiple comparisons. a Significantly different from the normal untreated group (Control), b Significantly different from the S. mansoni -infected untreated group (infected), c Significantly different from Inf + PZQ group, and d Significantly different from Inf + Vilda/DDS group, at P < 0.05. Inf + PZO, infected mice treated with paraziquantel; Inf + Vilda/DDS, infected mice treated with a combination of vildagliptin and diaminodiphenyl sulphone; Inf + Vilda, infected mice treated with vildagliptin; Inf + DDS, infected mice treated with diaminodiphenyl sulphone. All treatments were administered for 14 consecutive days, except PZQ for two consecutive days.

Article Snippet: Regarding immunohistochemical examination, thick deparaffinized liver Sects. (3–5 μm) were evaluated for the immunohistochemical reactivity of Anti-Rabbit α-SMA polyclonal Antibody (Cat#E-AB-34268, Elabscience, USA) and Anti-TGF-beta-1 polyclonal Antibody (Cat#ARG56429, Ari Gobio), following the manufacturer’s procedure.

Techniques: Infection, Immunohistochemical staining, Staining, Control, Expressing, Marker

Journal: Scientific reports

Article Title: An inspired microenvironment of cell replicas to induce stem cells into keratocyte-like dendritic cells for corneal regeneration.

doi: 10.1038/s41598-023-42359-9

Figure Lengend Snippet: Figure 2. Keratocytes and ADSCs characterization. (a) Light microscopy photographs of cultured keratocytes with dendritic morphology. (b) The gene expression profile demonstrated higher expression of positive keratocyte markers, including Lumican (LUM), Keratocan (KERA), ALDH3A1, and CD34 and minimal expression of myofibroblast marker (ACTA2) in keratocytes and ADSCs. (c,d) Immunofluorescence images of human keratocytes cultured for Keratocan (green)/lumican (red)/nuclei (DAPI, blue), and α-SMA (green)/ nuclei (DAPI, blue). (e) Representative of average protein levels. (f) Light inverted microscopy photographs of ADSCs with spindle shape morphology. (g) Flow cytometry results showing up-expression of CD105, CD73, CD90, and down-expression of CD45 in hADSCs.

Article Snippet: Cell 4 Vol:. (1234567890) Scientific Reports | (2023) 13:15012 | https://doi.org/10.1038/s41598-023-42359-9 nuclei were counterstained with 4,6-diamidino-2-phenylindole (DAPI) (1 mg/ mL; Santa Cruz Biotechnology, USA) for 5 min. To examine myofibroblast phenotype, the cells after fixation and permeabilization were diluted by rabbit polyclonal antibody to α-SMA (1:100 dilution, Biorbyt, UK) overnight at 4 °C.

Techniques: Light Microscopy, Cell Culture, Gene Expression, Expressing, Marker, Immunofluorescence, Inverted Microscopy, Flow Cytometry

Journal: Scientific reports

Article Title: An inspired microenvironment of cell replicas to induce stem cells into keratocyte-like dendritic cells for corneal regeneration.

doi: 10.1038/s41598-023-42359-9

Figure Lengend Snippet: Figure 8. Immunocytochemistry of seeded cells for protein expression after 2 and 3 weeks’ cultivation on patterned PDMS (KBM), patterned PDMS (DMEM/F12) and plain PDMS (KBM) at 40 X magnification. (a,d) Double staining for lumican (red) and keratocan (green) in ADSCs cultivated on patterned PDMS (KBM), patterned PDMS (DMEM/F12), and plain PDMS (KBM) after 14 and 21-days. (b,e) Expression of α-SMA protein (green) in the ADSCs on patterned PDMS (KBM and DMEM/F12), and plain PDMS (KBM). (c,f) Statistical analyses for cultured ADSCs on the imprinted substrates (KBM and DMEM/F12), compared to plain PDMS (KBM) at day 14 and 21, respectively. Data are mean ± SD. ***p < 0.001. ns nonsignificant.

Article Snippet: Cell 4 Vol:. (1234567890) Scientific Reports | (2023) 13:15012 | https://doi.org/10.1038/s41598-023-42359-9 nuclei were counterstained with 4,6-diamidino-2-phenylindole (DAPI) (1 mg/ mL; Santa Cruz Biotechnology, USA) for 5 min. To examine myofibroblast phenotype, the cells after fixation and permeabilization were diluted by rabbit polyclonal antibody to α-SMA (1:100 dilution, Biorbyt, UK) overnight at 4 °C.

Techniques: Immunocytochemistry, Expressing, Double Staining, Cell Culture

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