Journal: Drug Design, Development and Therapy

Article Title: Attenuation of everolimus-induced cytotoxicity by a protective autophagic pathway involving ERK activation in renal cell carcinoma cells

doi: 10.2147/DDDT.S160557

Figure Lengend Snippet: Everolimus (RAD001) induces autophagy in RCCs. Notes: ( A , B ) 786-O and A498 cell lines were treated by RAD001 with different concentrations (0, 0.01, 0.1, 1, 10, and 20 μmol/L) for 24 hours. ( A ) Immunoblotting showed that RAD001 induced autophagy indicators including upregulation of LC3-II/I and downregulation of p62 in a dose-dependent manner. ( B ) Quantification of the immunoblots. * P <0.05 versus 0 μmol/L group. n=3. ( C , D ) 786-O and A498 cell lines were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( C ) Immunoblotting showed that RAD001 induced autophagy in 786-O and A498 cells as time prolonged, which peaked at 8 hours. ( D ) Quantification of the immunoblots. * P <0.05 versus 0 hour group. n=3. Abbreviation: RCCs, renal cell carcinoma cells.

Article Snippet: The small molecular inhibitors RAD001 and AZD6244 were obtained from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Attenuation of everolimus-induced cytotoxicity by a protective autophagic pathway involving ERK activation in renal cell carcinoma cells

doi: 10.2147/DDDT.S160557

Figure Lengend Snippet: Inhibition of autophagy enhances RAD001-induced cytotoxicity, RAD001-induced activation of ERK signaling pathway in RCCs. Notes: 786-O and A498 cells were pretreated with CQ (autophagy inhibitor) (10 mmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( A ) MTT assay showed that RAD001 impaired cell viability of 786-O and A498 cells in a dose-dependent manner. n=3. ( B ) Immunoblotting of LC3-II/I and p62 showed that CQ could restore the downregulation of p62, but would not impair the upregulation of LC3-II/I because CQ blocks autophagy at the lysosomal degradation step. ( C ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control. # P <0.05 versus RAD001 group. n=3. ( D ) MTT assay displayed that autophagy inhibition by CQ promoted RAD001-induced cytotoxicity in 786-O and A498 cells. * P <0.05 versus control. # P <0.05 versus RAD001 group. n=3. ( E ) The apoptosis indicator cleave-PARP detected by immunoblot indicated that cells apoptosis was increased in 786-O and A498 cells after CQ intervention, compared to the RAD001 group. ( F ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. 786-O and A498 cells were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( G ) Immunoblotting analyzed the JNK, p38, and ERK signaling molecules in 786-O and A498 cells after RAD001 treatment. ( H ) Quantification of the immunoblots. * P <0.05 versus 0 hour group. n=3. Abbreviations: CQ, chloroquine; Con, control; RCCs, renal cell carcinoma cells; ERK, extracellular signal-regulated kinase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PARP, poly ADP ribose polymerase; JNK, c-Jun N-terminal kinase.

Article Snippet: The small molecular inhibitors RAD001 and AZD6244 were obtained from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Activation Assay, MTT Assay, Western Blot, Control

Journal: Drug Design, Development and Therapy

Article Title: Attenuation of everolimus-induced cytotoxicity by a protective autophagic pathway involving ERK activation in renal cell carcinoma cells

doi: 10.2147/DDDT.S160557

Figure Lengend Snippet: Selumetinib (AZD6244) significantly enhances RAD001-induced apoptosis of RCCs. Notes: 786-O and A498 cells were pretreated with AZD6244 (1 μmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( A ) Immunoblotting showed that AZD6244 significantly inhibited the activation of ERK pathway in 786-O and A498 cells. ( B ) Quantification of the immunoblots. ( C ) MTT assay detected the effect of AZD6244 on cell viability of 786-O and A498 cells, which suggested that AZD6244 effectively blunts RAD001-induced cell viability. ( D ) Immunoblotting demonstrates that AZD6244 could effectively restore upregulation of LC3-II/I and downregulation of p62 induced by RAD001 in 786-O and A498 cells. ( E ) Densitometric analysis was performed to quantify the immunoblots. ( F ) Immunoblot analysis indicated that combination treatment of RAD001 with AZD6244 could increase cleave-PARP level in 786-O and A498 cells, compared to the RAD001 group. ( G ) Densitometric analysis was performed to quantify the cleave-PARP immunoblots. ( H ) The difference of apoptosis determined by flow cytometry showed that AZD6244 significantly increased RAD001-induced cells apoptosis in 786-O and A498 cell lines. ( I ) Quantification of the apoptosis proportion. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. Abbreviations: RCCs, renal cell carcinoma cells; ERK, extracellular signal-regulated kinase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PARP, poly ADP ribose polymerase; FITC, fluorescein isothiocyanate; PI, propidium iodide; RAD001, everolimus; AZD6244, selumetinib; Con, control.

Article Snippet: The small molecular inhibitors RAD001 and AZD6244 were obtained from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: Western Blot, Activation Assay, MTT Assay, Flow Cytometry, Control

Journal: Drug Design, Development and Therapy

Article Title: Attenuation of everolimus-induced cytotoxicity by a protective autophagic pathway involving ERK activation in renal cell carcinoma cells

doi: 10.2147/DDDT.S160557

Figure Lengend Snippet: ERK mediated autophagy through upregulating Beclin-1 and p-Bcl-2 expression. Notes: ( A , B ) 786-O and A498 cells were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( A ) Immunoblotting showed that RAD001 induced upregulation of downstream signaling molecules of the ERK pathway including Beclin-1 and Bcl-2 in 786-O and A498 cells in a time-dependent manner. ( B ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus 0 hour group. n=3. ( C , D ) 786-O and A498 cells were pretreated with AZD6244 (1 μmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( C ) Immunoblotting showed that AZD6244 significantly decreased protein levels of Beclin-1 and Bcl-2 in 786-O and A498 cells. ( D ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. Abbreviations: ERK, extracellular signal-regulated kinase; RAD001, everolimus.

Article Snippet: The small molecular inhibitors RAD001 and AZD6244 were obtained from MedChem Express (Monmouth Junction, NJ, USA).

Techniques: Expressing, Western Blot, Control

Journal: Antioxidants & Redox Signaling

Article Title: Vascular Adhesion Protein-1: A Cell Surface Amine Oxidase in Translation

doi: 10.1089/ars.2017.7418

Figure Lengend Snippet: Crystallographic structure of VAP-1 . (A) Two identical monomers are colored blue and wheat . Copper (Cu) ion is orange and TPQ in each chain is presented as green spheres . (B) Docking of Siglec-9 peptide ( green ) into the active site of VAP-1. This binding mode is presented in Aalto et al. and assumes that the peptide binds covalently to TPQ. Courtesy of Dr. Tiina Salminen. Siglec, sialic acid-binding immunoglobulin-type lectins; TPQ, topaquinone; VAP-1, vascular adhesion protein-1.

Article Snippet: Astellas, Pharmaxis, and Promaxigen have developed small molecular inhibitors against VAP-1.

Techniques: Binding Assay

Journal: Antioxidants & Redox Signaling

Article Title: Vascular Adhesion Protein-1: A Cell Surface Amine Oxidase in Translation

doi: 10.1089/ars.2017.7418

Figure Lengend Snippet: VAP-1 is expressed in vascular endothelium and smooth muscle in human liver and tonsil. Triple stainings using anti-VAP-1, anti-smooth muscle actin (SMA), and anti-CD31 (pan-endothelial antibodies) are shown. Note that liver sinusoids ( arrowheads ) stain brightly with anti-VAP-1 but are devoid of smooth muscle present in larger vessels. Similarly, smaller capillaries are VAP-1/CD31 positive but lack SMA in tonsil ( arrows ).

Article Snippet: Astellas, Pharmaxis, and Promaxigen have developed small molecular inhibitors against VAP-1.

Techniques: Staining

Journal: Antioxidants & Redox Signaling

Article Title: Vascular Adhesion Protein-1: A Cell Surface Amine Oxidase in Translation

doi: 10.1089/ars.2017.7418

Figure Lengend Snippet: Functions of VAP-1 in health and disease. VAP-1 has multiple different physiological functions ( blue boxes ), and it is involved in their aberrations during different disease states ( yellow boxes ). Many of these processes are interdependent, and the role of VAP-1 in leukocyte extravasation is likely to contribute heavily to many of them.

Article Snippet: Astellas, Pharmaxis, and Promaxigen have developed small molecular inhibitors against VAP-1.

Techniques:

Journal: Antioxidants & Redox Signaling

Article Title: Vascular Adhesion Protein-1: A Cell Surface Amine Oxidase in Translation

doi: 10.1089/ars.2017.7418

Figure Lengend Snippet: A working model for VAP-1 function in the leukocyte extravasation cascade. A blood-borne leukocyte makes sequential contacts with the endothelial cell expressing VAP-1. When the two cell types (STEP 1) come in contact with each other (STEP 2) , a leukocyte counter-receptor of VAP-1 interacts in an enzyme activity-independent manner with the endothelial VAP-1. Thereafter, the same (or another) leukocyte surface molecule is used as a substrate in the VAP-1-mediated oxidative deamination reaction (STEP 3) . This results in the formation of a covalent but transient binding between the two cell types. After the catalytic reaction, the leukocyte surface molecule is modified into an aldehyde, a signaling molecule hydrogen peroxide is formed, and VAP-1 enzyme is converted back to the original state (STEP 4) . Note that the order of the proposed STEPs 2 and 3 is hypothetical.

Article Snippet: Astellas, Pharmaxis, and Promaxigen have developed small molecular inhibitors against VAP-1.

Techniques: Expressing, Activity Assay, Binding Assay, Modification

Journal: Antioxidants & Redox Signaling

Article Title: Vascular Adhesion Protein-1: A Cell Surface Amine Oxidase in Translation

doi: 10.1089/ars.2017.7418

Figure Lengend Snippet: VAP-1 can be used as a target for imaging. Representative sagittal ( left ), transaxial ( middle ), and coronal ( right ) multiplane PET images of [ 18 F]FDR-Siglec-9 (VAP-1 ligand) biodistribution in a rat. The images are summation from 10 to 60 min postinjection. Reproduced with permission from Chemical Communication [from Li et al. ].

Article Snippet: Astellas, Pharmaxis, and Promaxigen have developed small molecular inhibitors against VAP-1.

Techniques: Imaging

Journal: Frontiers in Pharmacology

Article Title: Berberine Attenuates Chronic Atrophic Gastritis Induced by MNNG and Its Potential Mechanism

doi: 10.3389/fphar.2021.644638

Figure Lengend Snippet: Primers sequences of real-time PCR analyses for mRNA expression.

Article Snippet: SB-431542 (Cat No. HY-10431, MedChem Express, Shanghai, China, 10 μΜ) , a small molecular inhibitor, was used to inhibit the expression of TGF-β1, and Rapamycin (Cat No. HY-10219,MedChem Express, Shanghai, China, 100 nΜ) ( ) was used to inhibit the expression of mTOR.

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: Frontiers in Pharmacology

Article Title: Berberine Attenuates Chronic Atrophic Gastritis Induced by MNNG and Its Potential Mechanism

doi: 10.3389/fphar.2021.644638

Figure Lengend Snippet: Antibodies.

Article Snippet: SB-431542 (Cat No. HY-10431, MedChem Express, Shanghai, China, 10 μΜ) , a small molecular inhibitor, was used to inhibit the expression of TGF-β1, and Rapamycin (Cat No. HY-10219,MedChem Express, Shanghai, China, 100 nΜ) ( ) was used to inhibit the expression of mTOR.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Berberine Attenuates Chronic Atrophic Gastritis Induced by MNNG and Its Potential Mechanism

doi: 10.3389/fphar.2021.644638

Figure Lengend Snippet: Histogram of TGF- β1 (A) , PI3K (B) , Akt (C) , mTOR (D) , LC3-II (E) and Beclin-1 (F) mRNA expression in MNNG co-cultured GES-1 cells by RT-PCR. Use SB-431542 and rapamycin to inhibit TGF- β1 and mTOR respectively. Data were shown as mean ± SD ( N = 4). **<0.01 vs. control group; *<0.05 vs. control group. ##<0.01 vs. MNNG group; #<0.05 vs. MNNG group.

Article Snippet: SB-431542 (Cat No. HY-10431, MedChem Express, Shanghai, China, 10 μΜ) , a small molecular inhibitor, was used to inhibit the expression of TGF-β1, and Rapamycin (Cat No. HY-10219,MedChem Express, Shanghai, China, 100 nΜ) ( ) was used to inhibit the expression of mTOR.

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Pharmacology

Article Title: Berberine Attenuates Chronic Atrophic Gastritis Induced by MNNG and Its Potential Mechanism

doi: 10.3389/fphar.2021.644638

Figure Lengend Snippet: Histogram of TGF- β1 (A) , PI3K (B) , P70S6K (C) , PTEN (D) , IL-6 (E) and IL-1β (F) mRNA expression in gastric tissue of CAG rats by RT-PCR. Data were shown as mean ± SD ( N = 5). **<0.01 vs. control group. ##<0.01 vs. MNNG group; #<0.05 vs. MNNG group. Ctrl, Control; BBR L, BBR low dose group; BBR H, BBR high dose group.

Article Snippet: SB-431542 (Cat No. HY-10431, MedChem Express, Shanghai, China, 10 μΜ) , a small molecular inhibitor, was used to inhibit the expression of TGF-β1, and Rapamycin (Cat No. HY-10219,MedChem Express, Shanghai, China, 100 nΜ) ( ) was used to inhibit the expression of mTOR.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Pharmacology

Article Title: Berberine Attenuates Chronic Atrophic Gastritis Induced by MNNG and Its Potential Mechanism

doi: 10.3389/fphar.2021.644638

Figure Lengend Snippet: Effect of BBR on proteins expression in CAG rats (A) Western blotting images of TGF-β1, PI3K, p-Akt, Akt, p-mTOR, mTOR (B) Relative TGF-β1 protein level in stomach (C) Relative PI3K protein level in stomach (D) Relative p-Akt/Akt protein level in stomach (E) Relative p-mTOR/mTOR protein level in stomach. Data were shown as mean ± SD ( N = 3). **<0.01 vs. control group; *<0.05 vs. control group. ##<0.01 vs. MNNG group; #<0.05 vs. MNNG group. Ctrl, Control; BBR L, BBR low dose group; BBR H, BBR high dose group.

Article Snippet: SB-431542 (Cat No. HY-10431, MedChem Express, Shanghai, China, 10 μΜ) , a small molecular inhibitor, was used to inhibit the expression of TGF-β1, and Rapamycin (Cat No. HY-10219,MedChem Express, Shanghai, China, 100 nΜ) ( ) was used to inhibit the expression of mTOR.

Techniques: Expressing, Western Blot

Journal: Molecular immunology

Article Title: TLR4 induced Wnt3a-Dvl3 restrains the intensity of inflammation and protects against endotoxin-driven organ failure through GSK3β/β-catenin signaling

doi: 10.1016/j.molimm.2019.12.013

Figure Lengend Snippet: Purified human monocytes and THP1 cells were pretreated with different Wnt3a inhibitors (100nM IWP-2 or 1μM PNU-74654) for 2 hours or transfected with pre-validated siRNA targeting Wnt3a or Dvl3 for 48 hours, then stimulated with E. coil LPS (1μg/ml). After 24 hours stimulation, the cell-free supernatants were collected and the production of IL-12P40, IL-6, and TNFα by monocytes (A) and THP1 (B) was measured by ELISA. (C) Silencing of wnt3a or dvl3 gene in human monocytes robustly reduces the expression of Wnt3a and Dvl3, but didn’t affect other proteins (Wnt7 or Dvl2). D and E, the production of IL-12P40, IL-6, and TNFα in LPS stimulated human monocytes with or without the pre-treatment of Wnt3a siRNA (C) or Dvl3 siRNA (E). The mRNA levels of inflammatory cytokines were measured by qRT-PCR in Dvl3 siRNA-treated monocytes after 3 hours stimulation with LPS (F). *, and *** indicates statistically significant at P<0.05, and P<0.001, respectively. Data represent the arithmetic mean±S.D. of three independent experiments.

Article Snippet: The small molecular inhibitors, N -(6-methyl-2-benzothiazolyl)-2-[(3,4,6,7-tetrahydro-4-oxo-3-phenylthieno[3,2- d ] pyrimidin-2-yl) thio]-acetamide (IWP-2) and benzoic acid, 2-phenoxy-,2-[(5-methyl-2-furanyl) methylene] hydrazide (PNU-74654), were from Tocris Biosciences (Minneapolis, MN).

Techniques: Purification, Transfection, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: bioRxiv

Article Title: PB-scope: Contrastive learning of dynamic processing body formation reveals undefined mechanisms of approved compounds

doi: 10.1101/2025.06.14.659731

Figure Lengend Snippet: MG132 induces a continuous reduction in P-bodies over 8 hours, while thapsigargin causes a temporary decrease in their number from 1 to 3 hours, followed by recovery after 4 hours. These findings are consistent with previous findings – .

Article Snippet: The library of 278 FDA-approved kinase inhibitor compounds and small molecular compounds MG132 and thapsigargin were purchased from TargetMol ( ).

Techniques:

Journal: Nature communications

Article Title: CDK9 recruits HUWE1 to degrade RARα and offers therapeutic opportunities for cutaneous T-cell lymphoma.

doi: 10.1038/s41467-024-54354-3

Figure Lengend Snippet: Fig. 1 | Chemical biology screening identifies a vulnerability to CDK9 inhibition of CTCL. a HH and Hut78 cells were treated with compounds (2 µM) from inhibitor library (L1200) and cell viability was evaluated using CCK8 assay after treatment for 48 hours. CDK Cyclin-dependent kinase, JAK/STAT Janus kinase-signal transducer and activator of transcription, PI3K Phosphoinositide 3-kinases, mTOR mammalian target of rapamycin complex. b The most effective kinase inhibitors (cell viabi- lity < 25%) identified from compound screening for HH and Hut78 cells. c Numbers of effective (red, cell viability < 25%) and ineffective (gray, cell viability ≥25%) CDK inhibitors from each different CDK subfamilies were shown. d IC50 curve of five representative CDK9 inhibitors, AT7519, Dinaciclib, Flavopiridol, SNS-032 and P276-00 in HH and Hut78 cells. Cell viability was counted by trypan blue staining. Results represent biologically independent experiments of n = 3. e–h Nude mice were subcutaneously injected with HH cells and randomly divided into Vehicle, Flavopiridol and SNS-032 groups (n = 8). Tumor volumes were measured at dif- ferent time points (e, g). At 17 days after subcutaneous injection, tumors were harvested and weighed (f, h). i Western blot analysis of indicated proteins from

Article Snippet: The small molecular inhibitors library (Selleckchem L1200) and the CDK inhibitors library (MedChemExpress, HYCPK9807, HYCPK9808, HYCPK9809, HYCPK9810) were applied.

Techniques: Inhibition, CCK-8 Assay, Staining, Injection, Western Blot