Journal: Oxidative Medicine and Cellular Longevity

Article Title: Elucidating the Novel Mechanism of Ligustrazine in Preventing Postoperative Peritoneal Adhesion Formation

doi: 10.1155/2022/9226022

Figure Lengend Snippet: Ligustrazine suppresses hypoxia-induced PMC functions by activating PPAR γ . (a, b) Western blot and qRT-PCR analysis of PPAR γ levels in hypoxia-induced PMCs treated with different concentrations of ligustrazine for 24 h. Compared with the control, ∗ P < 0.05 and ∗∗ P < 0.01. (c) Immunofluorescence analysis of PPAR γ nuclear distribution in hypoxia-induced PMCs treated with 20 μ M ligustrazine for 24 h (200x). (d) Flow cytometry analysis of PPAR γ nuclear translocation in hypoxia-induced PMCs treated with 20 μ M ligustrazine and/or 1 μ M GW9662 for 24 h. (e) Western blot analysis of protein expression of profibrotic cytokines (VEGF and CTGF) in hypoxia-induced PMCs treated with 20 μ M ligustrazine and/or 1 μ M GW9662 for 24 h. (f, g) FDA staining assay of cell viability in hypoxia-induced PMCs treated with 20 μ M ligustrazine and/or 1 μ M GW9662 for 24 h (200x). Compared with the control, ∗ P < 0.05 and ∗∗ P < 0.01. Compared with the ligustrazine group, # P < 0.05 and ## P < 0.01. (h) Western blot analysis of protein expression of migration and adhesion-associated molecules (VCAM-1 and ICAM-1) in hypoxia-induced PMCs treated with 20 μ M ligustrazine and/or 1 μ M GW9662 for 24 h. (i) Western blot analysis of cytoskeleton protein (vinculin) expression in hypoxia-induced PMCs treated with 20 μ M ligustrazine and/or 1 μ M GW9662 for 24 h. (j) Western blot analysis of protein expression of FSP1 in hypoxia-induced PMCs treated with 20 μ M ligustrazine and/or 1 μ M GW9662 for 24 h. (k) Western blot analysis of protein expression of ECM-associated molecules (MMP2 and TIMP-1), mesothelial-related phenotypic biomarkers (E-cadherin and cytokeratin 18), and mesenchymal-related phenotypic biomarkers (Snail and α -SMA) in hypoxia-induced PMCs treated with 20 μ M ligustrazine and/or 1 μ M GW9662 for 24 h.

Article Snippet: VEGF (sc-57496, Santa Cruz, USA), CTGF (sc-373936, Santa Cruz, USA), vascular cell adhesion molecule-1 (VCAM-1; ab134047, Abcam, USA), intercellular cell adhesion molecular-1 (ICAM-1; sc-8439, Santa Cruz, USA), cofilin (bs-2759R, Bioss, China), phospho-cofilin (p-cofilin, bs-20261R, Bioss, China), fibroblast-specific protein 1 (FSP1; bs-3759R, Bioss, China), matrix metalloproteinases 2 (MMP2; bs-4605R, Bioss, China), tissue inhibitor of metalloproteinases-1 (TIMP-1; bs-0415R, Bioss, China), E-cadherin (bs-1519R, Bioss, China), cytokeratin 18 (bs-2043R, Bioss, China), Snail (bs-1371R, Bioss, China), α -smooth muscle actin ( α -SMA; bs-10196R, Bioss, China), F-actin (FITC-conjugated phalloidin, C1033, Beyotime, China), vinculin (bs-6640R, Bioss, China), peroxisome proliferator-activated receptor γ (PPAR γ ; bs-4590R, Bioss, China), lamin B1 (sc-377000, Santa Cruz, USA), β -actin (CW0096M, CWBIO, China), and hypoxia-inducible factor 1 α (HIF-1 α ; sc-71247, Santa Cruz, USA) were used.

Techniques: Western Blot, Quantitative RT-PCR, Immunofluorescence, Flow Cytometry, Translocation Assay, Expressing, Staining, Migration

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Elucidating the Novel Mechanism of Ligustrazine in Preventing Postoperative Peritoneal Adhesion Formation

doi: 10.1155/2022/9226022

Figure Lengend Snippet: The suppression effect of ligustrazine is achieved by the activated PPAR γ on the transrepression of SMRT-mediated HIF-1 α . (a) Western blot analysis of protein expression of profibrotic cytokines (VEGF and CTGF) in hypoxia-induced PMCs treated with KC7F2 for 24 h. (b, c) FDA staining assay of cell viability in hypoxia-induced PMCs treated with KC7F2 for 24 h (200x). Compared with the control, ∗ P < 0.05 and ∗∗ P < 0.01. (d) Western blot analysis of protein expression of migration and adhesion-associated molecules (VCAM-1 and ICAM-1) in hypoxia-induced PMCs treated with KC7F2 for 24 h. (e) Western blot analysis of cytoskeleton protein (vinculin) expression in hypoxia-induced PMCs treated with KC7F2 for 24 h. (f) Western blot analysis of protein expression of FSP1 in hypoxia-induced PMCs treated with KC7F2 for 24 h. (g) Western blot analysis of protein expression of ECM-associated molecules (MMP2 and TIMP-1), mesothelial-related phenotypic biomarkers (E-cadherin and cytokeratin 18), and mesenchymal-related phenotypic biomarkers (Snail and α -SMA) in hypoxia-induced PMCs treated with KC7F2 for 24 h. (h, i) qRT-PCR and Western blot analysis of HIF-1 α levels in hypoxia-induced PMCs treated with 20 μ M ligustrazine and/or 1 μ M GW9662 for 24 h. Compared with the control, ∗ P < 0.05 and ∗∗ P < 0.01. Compared with the ligustrazine group, # P < 0.05 and ## P < 0.01. (j, k) qRT-PCR analysis of mRNA levels of SMRT and NCoR in hypoxia-induced HPMCs after different transfection for 24 h. Compared with the control, ∗ P < 0.05 and ∗∗ P < 0.01. (l) qRT-PCR analysis of mRNA levels of HIF-1 α in hypoxia-induced HPMCs treated with 20 μ M ligustrazine and/or with different transfection for 24 h. Compared with the control, ∗ P < 0.05 and ∗∗ P < 0.01. Compared with the ligustrazine group, # P < 0.05 and ## P < 0.01. (m) Western blot analysis of protein expression of HIF-1 α in hypoxia-induced HPMCs treated with 20 μ M ligustrazine and/or with different transfection for 24 h.

Article Snippet: VEGF (sc-57496, Santa Cruz, USA), CTGF (sc-373936, Santa Cruz, USA), vascular cell adhesion molecule-1 (VCAM-1; ab134047, Abcam, USA), intercellular cell adhesion molecular-1 (ICAM-1; sc-8439, Santa Cruz, USA), cofilin (bs-2759R, Bioss, China), phospho-cofilin (p-cofilin, bs-20261R, Bioss, China), fibroblast-specific protein 1 (FSP1; bs-3759R, Bioss, China), matrix metalloproteinases 2 (MMP2; bs-4605R, Bioss, China), tissue inhibitor of metalloproteinases-1 (TIMP-1; bs-0415R, Bioss, China), E-cadherin (bs-1519R, Bioss, China), cytokeratin 18 (bs-2043R, Bioss, China), Snail (bs-1371R, Bioss, China), α -smooth muscle actin ( α -SMA; bs-10196R, Bioss, China), F-actin (FITC-conjugated phalloidin, C1033, Beyotime, China), vinculin (bs-6640R, Bioss, China), peroxisome proliferator-activated receptor γ (PPAR γ ; bs-4590R, Bioss, China), lamin B1 (sc-377000, Santa Cruz, USA), β -actin (CW0096M, CWBIO, China), and hypoxia-inducible factor 1 α (HIF-1 α ; sc-71247, Santa Cruz, USA) were used.

Techniques: Western Blot, Expressing, Staining, Migration, Quantitative RT-PCR, Transfection

Journal: The EMBO Journal

Article Title: An EpCAM/Trop2 mechanostat differentially regulates collective behaviour of human carcinoma cells

doi: 10.1038/s44318-024-00309-9

Figure Lengend Snippet: Reagents and tools table

Article Snippet: vinculin , Boster Bio , MA1103 , Mouse , 1:100.

Techniques: Control, Recombinant, Plasmid Preparation, Modification, Western Blot, Electron Microscopy, Saline

Journal: Pharmacological Research - Modern Chinese Medicine

Article Title: The pharmaceutical applications of total flavonoids extract from Isatis tinctoria L. leaves

doi: 10.1016/j.prmcm.2022.100122

Figure Lengend Snippet: Fig. 8. Effects of TFE from Folium isatidis on the genes expression of TLR4 (a), TRAF6 (b), TRIF (c), I 𝜅B (d) and p65 (e) in lungs tissues of LPS induced ALI mice ( # compared with the control, ∗ compared with LPS, ∗ P < 0.05, ∗ ∗ / ## P < 0.01).

Article Snippet: Antibodies against TLR4, RAF6, TRIF, NF- κB p65, p-NF- κB p65, I κB α, p-I κB α and β-actin were urchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and econdary antibodies were obtained from Wuhan Boster Biological Techology., Ltd (Wuhan, China).

Techniques: Expressing, Control

Journal: Pharmacological Research - Modern Chinese Medicine

Article Title: The pharmaceutical applications of total flavonoids extract from Isatis tinctoria L. leaves

doi: 10.1016/j.prmcm.2022.100122

Figure Lengend Snippet: Fig. 9. Effects of TFE from Folium isatidis on the proteins expression of TLR4 (a), TRAF6 (b), TRIF (c), I 𝜅B (d), p-I 𝜅B (e), p65 (f) and p-p65 (g) in lungs tissues of LPS induced ALI mice ( # compared with the cntrol, ∗ compared with LPS, ∗ P < 0.05, ∗ ∗ / ## P < 0.01).

Article Snippet: Antibodies against TLR4, RAF6, TRIF, NF- κB p65, p-NF- κB p65, I κB α, p-I κB α and β-actin were urchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and econdary antibodies were obtained from Wuhan Boster Biological Techology., Ltd (Wuhan, China).

Techniques: Expressing

Journal: Pharmacological Research - Modern Chinese Medicine

Article Title: The pharmaceutical applications of total flavonoids extract from Isatis tinctoria L. leaves

doi: 10.1016/j.prmcm.2022.100122

Figure Lengend Snippet: Fig. 10. Regulation of TFE from Folium isatidis on TLR4-TRIF-NF- 𝜅B signaling pathway in LPS induced ALI mice.

Article Snippet: Antibodies against TLR4, RAF6, TRIF, NF- κB p65, p-NF- κB p65, I κB α, p-I κB α and β-actin were urchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and econdary antibodies were obtained from Wuhan Boster Biological Techology., Ltd (Wuhan, China).

Techniques:

Journal: Cell Reports

Article Title: SIN3A histone deacetylase action counteracts MUS81 to promote stalled fork stability

doi: 10.1016/j.celrep.2024.113778

Figure Lengend Snippet:

Article Snippet: Mouse anti Vinculin , Merk , Cat#V9264; RRID: AB_10603627.

Techniques: Recombinant, Control, Protease Inhibitor, Imaging, Reverse Transcription, Plasmid Preparation, Software, Magnetic Beads, Blocking Assay, In Situ, Western Blot, Membrane

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of CD147 expression alters cytoskeleton architecture and inhibits gelatinase production and SAPK pathway in human hepatocellular carcinoma cells

doi: 10.1186/1756-9966-27-50

Figure Lengend Snippet: Effects of si-CD147 on FAK and vinculin expression in SMMC-7721 cells . Total cellular protein from SMMC-7721 cells tranfected with siRNA was analyzed by 10% SDS-PAGE and transferred to PVDF membrane prior to immunoblotting. The PVDF membranes were washed and the bands were scanned by Odyssey Infrared Imaging System. The results showed that si-CD147 inhibited FAK and vinculiln expression in SMMC-7721 cells (A). The graph (B) compares scanning signal intensity of FAK and vinculin expression by ImageJ software.

Article Snippet: After being blocked with 5% (w/v) Odyssey blocking buffer (Li-COR Bioscience), the blots were incubated for overnight with antibodies directed against CD147 (BD Pharmingen), FAK(Sigma Aldrich, America) and vinculin (Merck Calbiochem) at 4°C.

Techniques: Expressing, SDS Page, Western Blot, Imaging, Software