rabbit  (Cell Signaling Technology Inc)

Journal: Pharmaceuticals

Article Title: Eupatorium lindleyanum DC Ameliorates Carbon Tetrachloride-Induced Hepatic Inflammation and Fibrotic Response in Mice

doi: 10.3390/ph18081228

Figure Lengend Snippet: Eup exerts anti-fibrotic effects by attenuating CCl 4 -induced liver fibrosis through modulation of the PDGF/PDGFR-β signaling pathway. ( A ) GSEA analyses of PDGF/PDGFR-β signaling pathway between the NC group and the 40 g/kg Eup group (n = 3 per group). ( B ) Representative immunoblot images demonstrating protein expression of GAPDH and PDGF-BB/PDGFR-β signaling pathway-related proteins. ( C – E ) The activation status of the PDGF-BB/PDGFR-β signaling pathway-related proteins was quantified through densitometric analysis using Image J (p-PDGFR-β/PDGFR-β, p-AKT/AKT, and p-ERK/ERK). GAPDH was the loading control. Data are presented as mean ± SD, n = 3. # p < 0.05 and ### p < 0.001 versus control group; * p < 0.05 and *** p < 0.001 versus CCl 4 group.

Article Snippet: For Western blot analysis, primary antibodies against β-actin (1:2500, abs171598, Absin, Shanghai, China), α-SMA (1:2500, Proteintech Group, Wuhan, China), Collagen I (1:2000, Proteintech Group, Wuhan, China), PDGFR-β (1:1250, Proteintech Group, Wuhan, China), p-PDGFR-β (1:1000, Abcam, UK), GAPDH (1:1500, Servicebio, Wuhan, China), p-AKT (1:1000, Abcam, UK), AKT (1:1000, Abcam, UK), p-ERK (1:1500, Selleck Chemicals, Houston, TX, USA) and ERK (1:1000, Selleck Chemicals, Houston, TX, USA), were applied.

Techniques: Western Blot, Expressing, Activation Assay, Control

Journal: Pharmaceuticals

Article Title: Eupatorium lindleyanum DC Ameliorates Carbon Tetrachloride-Induced Hepatic Inflammation and Fibrotic Response in Mice

doi: 10.3390/ph18081228

Figure Lengend Snippet: Eup suppresses PDGF-BB-induced HSC activation by inhibiting the PDGF-BB/PDGFR-β signaling pathway. ( A ) LX-2 cells were cultured with 20 ng/mL PDGF-BB, 1, 10, and 20 μg/mL Eup for 24 h. ( B – E ) qRT-PCR analysis of α-SMA , Col1 , Col3 , and LOX in LX-2 cells administrated with different concentrations of Eup. ( F ) Representative immunoblot images demonstrating protein expression of GAPDH, α-SMA, p-PDGFR-β, PDGFR-β, p-AKT, AKT, p-ERK, and ERK. ( G – J ) Expression levels of α-SMA and phosphorylation ratios of PDGFR-β (p-PDGFR-β/PDGFR-β), AKT (p-AKT/AKT), and ERK (p-ERK/ERK). The loading control was GAPDH. Data are presented as mean ± SD, n = 3. # p < 0.05, ## p < 0.01, and ### p < 0.001 versus PDGF-BB (-) Eup (-) group; * p < 0.05 and ** p < 0.01 versus PDGF-BB (+) Eup (-) group.

Article Snippet: For Western blot analysis, primary antibodies against β-actin (1:2500, abs171598, Absin, Shanghai, China), α-SMA (1:2500, Proteintech Group, Wuhan, China), Collagen I (1:2000, Proteintech Group, Wuhan, China), PDGFR-β (1:1250, Proteintech Group, Wuhan, China), p-PDGFR-β (1:1000, Abcam, UK), GAPDH (1:1500, Servicebio, Wuhan, China), p-AKT (1:1000, Abcam, UK), AKT (1:1000, Abcam, UK), p-ERK (1:1500, Selleck Chemicals, Houston, TX, USA) and ERK (1:1000, Selleck Chemicals, Houston, TX, USA), were applied.

Techniques: Activation Assay, Cell Culture, Quantitative RT-PCR, Western Blot, Expressing, Phospho-proteomics, Control

Journal: Pharmaceuticals

Article Title: Eupatorium lindleyanum DC Ameliorates Carbon Tetrachloride-Induced Hepatic Inflammation and Fibrotic Response in Mice

doi: 10.3390/ph18081228

Figure Lengend Snippet: The mechanism diagram of Eup in improving CCl 4 -induced liver fibrosis. Eup suppressed the PDGF-BB/PDGFR-β signaling pathway, thereby inhibiting HSCs activation, and then improving liver fibrosis induced by CCl 4 .

Article Snippet: For Western blot analysis, primary antibodies against β-actin (1:2500, abs171598, Absin, Shanghai, China), α-SMA (1:2500, Proteintech Group, Wuhan, China), Collagen I (1:2000, Proteintech Group, Wuhan, China), PDGFR-β (1:1250, Proteintech Group, Wuhan, China), p-PDGFR-β (1:1000, Abcam, UK), GAPDH (1:1500, Servicebio, Wuhan, China), p-AKT (1:1000, Abcam, UK), AKT (1:1000, Abcam, UK), p-ERK (1:1500, Selleck Chemicals, Houston, TX, USA) and ERK (1:1000, Selleck Chemicals, Houston, TX, USA), were applied.

Techniques: Activation Assay

Journal: Cancer Medicine

Article Title: PDGF ‐D Promotes Epithelial–Mesenchymal Transition of Glioma Cells Through the NF ‐ κB / NOTCH1 Pathway

doi: 10.1002/cam4.71002

Figure Lengend Snippet: PDGF‐D was highly expressed in glioma and promoted proliferation. (A) CGGA data showing the correlation between PDGF‐D expression in glioma and the degree of malignancy and prognosis. (B, C) Representative immunostaining images showing in situ expression of PDGF‐D and PDGFR‐β in glioma tissues and normal brain tissues. The brown‐stained regions correspond to positive staining. Images are 400× magnified. (D) PDGF‐D and PDGFR‐β protein expression in the indicated glioma cell lines. (E, G) Changes in PDGF‐D mRNA and protein expression after stable transfection of PDGF‐D sh1 and PDGF‐D sh2 in LN18 cells. (F, H) Changes in PDGF‐D mRNA and protein expression levels in U87 cells overexpressing PDGF‐D. (I) Colonies formed by the control and PDGF‐D‐knockdown LN18 cells. (J) Colonies formed by the control and PDGF‐D‐overexpressing U87 cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The antibodies for PDGFR‐β (#3169), MMP‐2 (#40994), N‐cadherin (#13116), E‐cadherin (#3195), Slug (#9585), β‐catenin (#8480), vimentin (#5741), NICD (#3608), and Hairy and Enhancement of Split 1 (Hes1) (#11988) were purchased from CST and were diluted 1:1000 for western blotting and 1:200 for IHC.

Techniques: Expressing, Immunostaining, In Situ, Staining, Stable Transfection, Control, Knockdown