Journal: Renal Failure

Article Title: Protective effect of phosphoenolpyruvate carboxykinase 1 on inflammation and fibrotic progression of IgA nephropathy

doi: 10.1080/0886022X.2025.2508297

Figure Lengend Snippet: The expression of PCK1 in renal tissue exhibited a negative correlation with the expression of inflammatory chemokines and fibrosis-related markers. Immunohistochemistry and quantification of TNF-α and IL-1β in human renal tissue (patients with IgAN, n = 79 and NC, n = 30). (B) Pearson correlation analysis of the relationship between PCK1 and TNF-α/IL-1β expression. Decreased PCK1 expression was linearly negatively correlated with TNF-α/IL-1β levels (r = -0.625/r = -0.607, p < 0.001); (C) Immunohistochemistry and quantification of TGF-β1, psmad3, α-SMA, FN, and col-IV in human renal tissue (patients with IgAN, n = 79 and NC, n = 30). (D) Pearson correlation analysis of the relationship between PCK1 and TGF-β1/psmad3/α-SMA/FN/col-IV expression. Decreased PCK1 expression was linearly negatively correlated with TGF-β1/psmad3/α-SMA/FN/col-IV levels (r = -0.505/r = -0.720/r = -0.613/r = -0.731/r = -0.668). Abbreviations: NC, normal control; TNF-α, tumor necrosis factor α; IL-1β, interleukin 1β; TGF-β1, transforming growth factor-β1; psmad3, Phosphorylated-smad3; α-SMA, α-smooth muscle actin; FN, fibronectin; col-IV, collagen-IV. *** p < 0.001.

Article Snippet: PPAR-γ antibody (16643-1-AP), PCK1 antibody (16754-1-AP), and fibronectin polyclonal antibody (15613-1-AP) were sourced from Proteintech Group (Wuhan, China).

Techniques: Expressing, Immunohistochemistry, Control

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Dioscin ameliorates peritoneal fibrosis by inhibiting epithelial-to-mesenchymal transition of human peritoneal mesothelial cells via the TLR4/MyD88/NF-κB signaling pathway

doi:

Figure Lengend Snippet: Dioscin inhibits LPS-induced fibrosis in HMrSV5 cells. Western blotting for detecting protein levels of α-SMA, collagen I and fibronectin. ###P < 0.001 compared with negative control group or dioscin (1.0 μg/ml) group. ***P < 0.001 compared with LPS group. NC, negative control; LPS, lipopolysaccharide; Dio, Dioscin; α-SMA, α-smooth muscle actin.

Article Snippet: Then the membranes were incubated with following primary antibodies: α-SMA (1:100, Proteintech Group), collagen I (1:100, Proteintech Group), fibronectin (1:250, Abcam), TLR4 (1:1000, Proteintech Group), MyD88 (1:1000, Proteintech Group), NF-κB (1:1000, Proteintech Group), TGF-β1 (1:1000, Proteintech Group), p-Smad2/Smad2 (1:1000, Proteintech Group), respectively.

Techniques: Western Blot, Negative Control

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Dioscin ameliorates peritoneal fibrosis by inhibiting epithelial-to-mesenchymal transition of human peritoneal mesothelial cells via the TLR4/MyD88/NF-κB signaling pathway

doi:

Figure Lengend Snippet: Dioscin inhibits EMT and fibrosis through TLR4/MyD88/NF-κB pathway in HMrSV5 cells. A. Western blotting for assessing protein levels of TLR4, MyD88, NF-κB, TGF-β1, p-Smad2, Smad2, α-SMA, collagen I and fibronectin. B. immunofluorescence assay for detecting expressions of α-SMA, collagen I and fibronectin in LPS+TLR4 group and LPS+TLR4+dioscin (0.5 μg/ml) group. ###P < 0.001 compared with negative control group or Ppicza group. *P < 0.05, **P < 0.01 and ***P < 0.001 compared with LPS+ pPICZA group or LPS+TLR4 group. NC, negative control; LPS, lipopolysaccharide; Dio, Dioscin; α-SMA, α-smooth muscle actin; MyD88, myeloid differentiation factor 88; NF-κB, nuclear factor κB; TGF-β1, transforming growth factor-β1; TLR4, Toll-like receptor (TLR) 4.

Article Snippet: Then the membranes were incubated with following primary antibodies: α-SMA (1:100, Proteintech Group), collagen I (1:100, Proteintech Group), fibronectin (1:250, Abcam), TLR4 (1:1000, Proteintech Group), MyD88 (1:1000, Proteintech Group), NF-κB (1:1000, Proteintech Group), TGF-β1 (1:1000, Proteintech Group), p-Smad2/Smad2 (1:1000, Proteintech Group), respectively.

Techniques: Western Blot, Immunofluorescence, Negative Control

Journal: Oncology Letters

Article Title: MicroRNA-432 is downregulated in cervical cancer and directly targets FN1 to inhibit cell proliferation and invasion

doi: 10.3892/ol.2019.10403

Figure Lengend Snippet: FN1 is a direct target gene of miR-432 in cervical cancer cells. (A) The WT binding sequences of miR-432 in the 3′-UTR of FN1 as predicted by a bioinformatics analysis. The MUT binding site was also generated in the seed region of the 3′-UTR of FN1. (B) pMIR-WT-FN1-3′-UTR or pMIR-MUT-FN1-3′-UTR was co-transfected with miR-432 mimic or miR-NC into HeLa and SiHa cells. Cells were collected and used for the detection of luciferase activity 48 h after transfection. (C) RT-qPCR was performed to determine the expression level of FN1 mRNA in cervical cancer tissues and matched adjacent normal tissues. (D) Spearman's correlation analysis was performed to investigate the correlation between miR-432 and FN1 mRNA levels in cervical cancer tissues. r=−0.5383, P<0.0001 (E) FN1 mRNA expression in HeLa and SiHa cells following transfection with miR-432 mimic or miR-NC was measured by RT-qPCR and western blot analysis. (F) Protein expression in HeLa and SiHa cells following transfection with miR-432 mimic or miR-NC was measured using western blot analysis. *P<0.05 vs. miR-NC; **P<0.05 vs. adjacent normal tissues. FN1, fibronectin 1; miR-432, microRNA-432; WT, wild-type; UTR, untranslated region; MUT, mutated; NC, negative control; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: Following blocking with 5% skimmed milk suspended in Tris-buffered saline-containing Tween-20 (TBST) at room temperature for 2 h, the membranes were incubated overnight at 4°C with primary antibodies as follows: Rabbit anti-human FN1 antibody (catalogue no. 15613-1-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and rabbit anti-human GAPDH antibody (catalogue no. ab181603; 1:1,000 dilution; Abcam, Cambridge, UK).

Techniques: Binding Assay, Generated, Transfection, Luciferase, Activity Assay, Quantitative RT-PCR, Expressing, Western Blot, Negative Control, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: MicroRNA-432 is downregulated in cervical cancer and directly targets FN1 to inhibit cell proliferation and invasion

doi: 10.3892/ol.2019.10403

Figure Lengend Snippet: FN1 silencing suppresses the cellular proliferative and invasive capacities of HeLa and CaSki cells. (A) FN1 protein level in HeLa and SiHa cells treated with FN1 siRNA or NC siRNA was detected using the reverse transcription-quantitative polymerase chain reaction. (B) Cell Counting Kit-8 was used to assess the proliferation of HeLa and SiHa cells upon FN1 inhibition. (C) Transwell Matrigel invasion assay was used to assess the invasion of HeLa and SiHa cells upon FN1 inhibition. *P<0.05 vs. NC siRNA. FN1, fibronectin 1; siRNA, small interfering RNA; NC, negative control; OD, optical density.

Article Snippet: Following blocking with 5% skimmed milk suspended in Tris-buffered saline-containing Tween-20 (TBST) at room temperature for 2 h, the membranes were incubated overnight at 4°C with primary antibodies as follows: Rabbit anti-human FN1 antibody (catalogue no. 15613-1-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and rabbit anti-human GAPDH antibody (catalogue no. ab181603; 1:1,000 dilution; Abcam, Cambridge, UK).

Techniques: Real-time Polymerase Chain Reaction, Cell Counting, Inhibition, Invasion Assay, Small Interfering RNA, Negative Control

Journal: Oncology Letters

Article Title: MicroRNA-432 is downregulated in cervical cancer and directly targets FN1 to inhibit cell proliferation and invasion

doi: 10.3892/ol.2019.10403

Figure Lengend Snippet: Recovered FN1 expression attenuates the inhibitory effects of miR-432 overexpression in the proliferation and invasion of HeLa and SiHa cells. miR-432 mimic along with FN1 overexpression plasmid pcDNA3.1-FN1 or pcDNA3.1 was introduced into HeLa and SiHa cells. (A) Western blot analysis was performed to determine the expression of FN1 protein. (B and C) The proliferation of the aforementioned cells was determined using a Cell Counting Kit-8. (C) The invasion of the aforementioned cells was determined by performing a Transwell Matrigel invasion assay. *P<0.05 vs. miR-NC; #P<0.05 vs. miR-432 mimics+pcDNA3.1. FN1, fibronectin 1; miR-432, microRNA-432; OD, optical density.

Article Snippet: Following blocking with 5% skimmed milk suspended in Tris-buffered saline-containing Tween-20 (TBST) at room temperature for 2 h, the membranes were incubated overnight at 4°C with primary antibodies as follows: Rabbit anti-human FN1 antibody (catalogue no. 15613-1-AP; 1:200 dilution; ProteinTech Group, Inc., Chicago, IL, USA) and rabbit anti-human GAPDH antibody (catalogue no. ab181603; 1:1,000 dilution; Abcam, Cambridge, UK).

Techniques: Expressing, Over Expression, Plasmid Preparation, Western Blot, Cell Counting, Invasion Assay