fluorogenic substrate mcaala pro lys dnp oh  (MedChemExpress)

Journal: World Journal of Gastroenterology

Article Title: Wogonin derivative V8 enhances bortezomib efficacy in gastric carcinoma by disrupting lysosome-mediated drug resistance

doi: 10.3748/wjg.v32.i8.113299

Figure Lengend Snippet: V8 synergizes with bortezomib to inhibit gastric cancer cell growth. A: Cell viability assay was performed in different gastric cancer cells treated with V8 and bortezomib (BTZ) for 24 hours; B: Combination index was calculated in different gastric cancer cells; C and D: MGC-803 cells were treated with V8 (9 μM) and BTZ (100 nM) for 24 hours. Western blotting was carried out for analysis of indicated protein expression with β-actin as loading control; E and F: Cells were treated with V8 (9 μM), BTZ (100 nM), or benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone (20 μM) for 24 hours, followed by flow cytometry assay for apoptosis analysis. Representative images (E) and quantitative data (F) are shown. b P < 0.01. d P < 0.0001. CI: Combination index; Bip: Immunoglobulin heavy chain binding protein; p-eIF2α: Phosphorylated eukaryotic translation initiation factor 2 alpha; Clpp: Caseinolytic mitochondrial matrix peptidase proteolytic subunit; DMSO: Dimethyl sulfoxide; BTZ: Bortezomib; V-FITC: Annexin V-fluorescein isothiocyanate; PI: Propidium iodide; Z-VAD-FMK: Benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone.

Article Snippet: BTZ (HY-10227), benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone (Z-VAD-FMK) (HY-16658B), chloroquine (HY-17589A), 3-methyladenine (3-MA) (HY-19312), bafilomycin A1 (Baf-A1) (HY-100558) and L-leucyl-L-leucine methyl ester hydrochloride (HY-129905) were purchased from MedChemExpress (Shanghai, China).

Techniques: Viability Assay, Western Blot, Expressing, Control, Flow Cytometry, Binding Assay

Journal: World Journal of Gastroenterology

Article Title: Wogonin derivative V8 enhances bortezomib efficacy in gastric carcinoma by disrupting lysosome-mediated drug resistance

doi: 10.3748/wjg.v32.i8.113299

Figure Lengend Snippet: Schematic diagram illustrating the mechanism by which V8 synergizes with bortezomib to induce cell death in gastric cancer cells. Bortezomib (BTZ) alone is sequestered in lysosomes, resulting in insufficient proteasome inhibition and mild apoptosis. V8 alone induces lysosomal damage and cell death via lysosomal membrane permeabilization and CTSB release. V8 disrupts lysosomes to release sequestered BTZ, enabling potent proteasome inhibition that triggers proteotoxic stress (ubiquitinated protein accumulation, endoplasmic reticulum stress, and mitochondrial stress), ultimately culminating in caspase-dependent apoptosis. BTZ: Bortezomib; TFEB: Transcription factor EB; CLEAR: Coordinated lysosomal expression and regulation; Bip: Immunoglobulin heavy chain binding protein; p-eIF2α: Phosphorylated eukaryotic translation initiation factor 2 alpha; Z-VAD-FMK: Benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone.

Article Snippet: BTZ (HY-10227), benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone (Z-VAD-FMK) (HY-16658B), chloroquine (HY-17589A), 3-methyladenine (3-MA) (HY-19312), bafilomycin A1 (Baf-A1) (HY-100558) and L-leucyl-L-leucine methyl ester hydrochloride (HY-129905) were purchased from MedChemExpress (Shanghai, China).

Techniques: Inhibition, Membrane, Expressing, Binding Assay

Journal: Infection and Immunity

Article Title: Chlamydia trachomatis restricts signaling through NOD2 until late in the pathogen’s developmental cycle

doi: 10.1128/iai.00472-25

Figure Lengend Snippet: Pre-treatment with NOD1- and NOD2-stimulatory ligands prior to infection with C. trachomatis significantly impacts inclusion size in NLR-expressing cell lines. HepG2, C-33A, and SiHa cells were pre-treated with Tri-DAP, MDP, or both, prior to infection with C. trachomatis L2 strain Bu/434. At 24 hpi, cells were fixed, inclusions were labeled, counted, and measured. zStacks of MOMP-labeled objects were obtained from a Zeiss 700 confocal microscope, and approximate volume measurements were calculated for all inclusions present within 10 imaging fields. Data presented represent all MOMP-labeled objects counted resulting in inclusions > 20 µm 3 ; 3 µm across with red lines indicating the mean of each group within a data set. Significance was assessed via one-way ANOVA with multiple comparisons. ****; P < 0.0001, ***; P < 0.001, **; P < 0.01, ns; not significant.

Article Snippet: MDP and L-Ala-γ-D-Glu-mDAP (Tri-DAP) were purchased from InvivoGen.

Techniques: Infection, Expressing, Labeling, Microscopy, Imaging

Journal: Infection and Immunity

Article Title: Chlamydia trachomatis restricts signaling through NOD2 until late in the pathogen’s developmental cycle

doi: 10.1128/iai.00472-25

Figure Lengend Snippet: Pre-treatment with NOD2-stimulatory ligands delays the development of C. trachomatis in NLR-expressing cell lines. HepG2 ( A ), C-33A ( B ), and SiHa ( C ) cells were either left untreated or pre-treated with the NOD1-stimulatory ligand Tri-DAP, the NOD2-stimulatory ligand MDP, or both Tri-DAP and MDP. After 24 hours, cells were infected with C. trachomatis , and the development of infectious EBs was assessed at the indicated time points (30, 48, and 72 hpi). Lines delineate the mean of three separate biological replicates with each replicate displayed as a colored dot. Significance was assessed via one-way ANOVA with multiple comparisons. **; P < 0.01, *; P < 0.05, ns; not significant.

Article Snippet: MDP and L-Ala-γ-D-Glu-mDAP (Tri-DAP) were purchased from InvivoGen.

Techniques: Expressing, Infection