anti col 1 α1  (Bioss)

Journal: Frontiers in Immunology

Article Title: A Novel miRNA From Egg-Derived Exosomes of Schistosoma japonicum Promotes Liver Fibrosis in Murine Schistosomiasis

doi: 10.3389/fimmu.2022.860807

Figure Lengend Snippet: Information for immunofluorescence antibodies and other reagents.

Article Snippet: Primary antibodies were anti-TGF-β RI (bs-0638R, Bioss, China), anti-GAPDH (5174S, Cell Signaling Technology, CST, USA), anti-α-SMA (19245S, CST), anti-Col 1 α1 (bs-7158R, Bioss), anti-Col 3 α1 (22734-1-AP, Proteintech, USA), anti-Smad3 (9523S, CST), anti-phospho-Smad3 (9520S, CST), anti-Flotillin-1 (3253S, CST), and anti-HSP70 (46477S, CST).

Techniques: Immunofluorescence

Journal: Frontiers in Immunology

Article Title: A Novel miRNA From Egg-Derived Exosomes of Schistosoma japonicum Promotes Liver Fibrosis in Murine Schistosomiasis

doi: 10.3389/fimmu.2022.860807

Figure Lengend Snippet: Primer sequences.

Article Snippet: Primary antibodies were anti-TGF-β RI (bs-0638R, Bioss, China), anti-GAPDH (5174S, Cell Signaling Technology, CST, USA), anti-α-SMA (19245S, CST), anti-Col 1 α1 (bs-7158R, Bioss), anti-Col 3 α1 (22734-1-AP, Proteintech, USA), anti-Smad3 (9523S, CST), anti-phospho-Smad3 (9520S, CST), anti-Flotillin-1 (3253S, CST), and anti-HSP70 (46477S, CST).

Techniques: Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: A Novel miRNA From Egg-Derived Exosomes of Schistosoma japonicum Promotes Liver Fibrosis in Murine Schistosomiasis

doi: 10.3389/fimmu.2022.860807

Figure Lengend Snippet: Egg-derived exosomes from Schistosoma japonicum can promote liver fibrosis. (A) Western blotting detection of markers of egg-derived exosomes of S. japonicum . (B) Transmission electron microscopy (TEM) analysis of exocrine. (C) At 24 h after stimulation of LX-2 human hepatic stellate) cells (HSCs) by egg-derived exosomes of S. japonicum , mRNA levels of smooth muscle actin (α-SMA), collagen (Col) 1 (α1) , and collagen (Col) 3 (α1) were measured by qPCR. Results are averaged from three independent experiments, and Student’s t -tests were applied to assess differences between two groups. The mRNA expression levels of α-SMA , Col 1 (α1) and Col 3 (α1) were normalized against GAPDH ** p < 0.001, *** p < 0.0005.

Article Snippet: Primary antibodies were anti-TGF-β RI (bs-0638R, Bioss, China), anti-GAPDH (5174S, Cell Signaling Technology, CST, USA), anti-α-SMA (19245S, CST), anti-Col 1 α1 (bs-7158R, Bioss), anti-Col 3 α1 (22734-1-AP, Proteintech, USA), anti-Smad3 (9523S, CST), anti-phospho-Smad3 (9520S, CST), anti-Flotillin-1 (3253S, CST), and anti-HSP70 (46477S, CST).

Techniques: Derivative Assay, Western Blot, Transmission Assay, Electron Microscopy, Expressing

Journal: Frontiers in Immunology

Article Title: A Novel miRNA From Egg-Derived Exosomes of Schistosoma japonicum Promotes Liver Fibrosis in Murine Schistosomiasis

doi: 10.3389/fimmu.2022.860807

Figure Lengend Snippet: Novel miRNA-33 mimics can activate HSCs (human hepatic stellate cells). (A) LX-2 cells were transfected with several novel miRNA mimics (novel miRNA-30, novel miRNA-33, novel miRNA-68, miR-1, miRNA-NC), cultivated for 24 h, and mRNA levels of α-SMA and Col 1 (α1) were measured by qPCR. (B) LX-2 cells were transfected with several novel miRNA mimics (novel miRNA-30, novel miRNA-33, novel miRNA-68, miR-1, miRNA-NC), cultivated for 48 h, and expression levels of α-SMA and Col 1 (α1) were measured by western blotting. Results were compared with those of the NC mimic group. The abundances of α-SMA and Col 1 (α1) proteins were normalized against GAPDH. Results are averaged from three independent experiments, and Student’s t -tests were applied to assess differences between two groups (** p < 0.001, **** p < 0.0001).

Article Snippet: Primary antibodies were anti-TGF-β RI (bs-0638R, Bioss, China), anti-GAPDH (5174S, Cell Signaling Technology, CST, USA), anti-α-SMA (19245S, CST), anti-Col 1 α1 (bs-7158R, Bioss), anti-Col 3 α1 (22734-1-AP, Proteintech, USA), anti-Smad3 (9523S, CST), anti-phospho-Smad3 (9520S, CST), anti-Flotillin-1 (3253S, CST), and anti-HSP70 (46477S, CST).

Techniques: Transfection, Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: A Novel miRNA From Egg-Derived Exosomes of Schistosoma japonicum Promotes Liver Fibrosis in Murine Schistosomiasis

doi: 10.3389/fimmu.2022.860807

Figure Lengend Snippet: Novel miRNA-33 agomir causes liver fibrosis in mice. Six-week-old female C57BL/6 mice were injected with agomir NC (Ribo), agomir novel 33 (Ribo), or normal saline once a week for 6 weeks. Results were compared with the agomir NC group; the normal group was only used as a reference without comparative analysis. Only the gray values of the agomir NC group and the agomir novel 33 group were statistically analyzed, the blank group was not analyzed. (A) Novel miRNA-33 levels in mouse liver and serum were increased significantly. (B) qPCR results showing upregulation of α-SMA, Col 1 (α1) , and Col 3 (α1) at the mRNA level. (C) Western blotting results showing upregulation of α-SMA, Col 1 (α1), and Col 3 (α1) at the protein level. The abundances of α-SMA, Col 1 (α1), and Col 3 (α1) proteins were normalized against GAPDH, and miRNA expression levels were normalized against U6. Results are averaged from three independent experiments, and Student’s t -tests were applied to assess differences between two groups (** p < 0.001, *** p < 0.0005, **** p < 0.0001).

Article Snippet: Primary antibodies were anti-TGF-β RI (bs-0638R, Bioss, China), anti-GAPDH (5174S, Cell Signaling Technology, CST, USA), anti-α-SMA (19245S, CST), anti-Col 1 α1 (bs-7158R, Bioss), anti-Col 3 α1 (22734-1-AP, Proteintech, USA), anti-Smad3 (9523S, CST), anti-phospho-Smad3 (9520S, CST), anti-Flotillin-1 (3253S, CST), and anti-HSP70 (46477S, CST).

Techniques: Injection, Western Blot, Expressing

Journal: Frontiers in Immunology

Article Title: A Novel miRNA From Egg-Derived Exosomes of Schistosoma japonicum Promotes Liver Fibrosis in Murine Schistosomiasis

doi: 10.3389/fimmu.2022.860807

Figure Lengend Snippet: Inhibition of novel miRNA-33 reduces the degree of liver fibrosis. Six-week-old female C57BL/6 mice were percutaneously exposed to 20 ± 1 S. japonicum cercariae, and 120 μL of 20 nM miRNA antagomir NC(Ribo), miRNA antagomir novel 33 (Ribo), or normal saline were injected via the tail vein 7 days after mice were infected with S. japonicum cercariae, once a week for 6 weeks. The magnification of Masson-stained images is 10 ×, and the magnification of immunofluorescence image is 20 ×. (A, B) Masson staining showing that areas of collagen were decreased after treatment with novel miRNA-33 antagomir. (C) Egg count in mouse liver tissue showing there was no statistical difference between the three groups. (D) Expression of novel miRNA-33 in mice infected with S. japonicum for 6 weeks. (E) qPCR results showing downregulation of α-SMA, Col 1 (α1), and Col 3 (α1) at the mRNA level. (F) Immunofluorescence images showing downregulation of type I collagen and α-SMA after treatment with antagomir. mRNA expression levels of α-SMA , Col 1 (α1) , and Col 3 (α1) were normalized agaisnt GAPDH , and expression of novel miRNA-33 was normalized against U6. Results are averaged from three independent experiments, Student’s t -tests were applied to assess differences between two groups, and one-way analysis of variance (ANOVA) followed by Tukey’s post-tests was used for three or more groups (fluorescence is SpGreen; n = 20; * p < 0.05, ** p < 0.001, *** p < 0.0005; NS, not significant).

Article Snippet: Primary antibodies were anti-TGF-β RI (bs-0638R, Bioss, China), anti-GAPDH (5174S, Cell Signaling Technology, CST, USA), anti-α-SMA (19245S, CST), anti-Col 1 α1 (bs-7158R, Bioss), anti-Col 3 α1 (22734-1-AP, Proteintech, USA), anti-Smad3 (9523S, CST), anti-phospho-Smad3 (9520S, CST), anti-Flotillin-1 (3253S, CST), and anti-HSP70 (46477S, CST).

Techniques: Inhibition, Injection, Infection, Staining, Immunofluorescence, Expressing, Fluorescence