dynamin inhibitor dynasore  (MedChemExpress)

Journal: bioRxiv

Article Title: GM-CSF and M-CSF Driven Differentiation Differentially Regulates Chikungunya Virus Infection and Antiviral Responses in Human Monocyte-Derived Macrophages

doi: 10.64898/2026.03.11.710213

Figure Lengend Snippet: (A) Schematic representation of compounds targeting CHIKV entry and internalization mechanisms in macrophages. GM-Mϕ cultured on 96-wells plates were incubated with macrophage media containing compounds inhibiting phagocytosis: (B) cytochalasin D, macropinocytosis: (C) EIPA, clathrin: (D) pitstop2, dynamin: (E) dynasore, or endocytosis/viral fusion: (F) bafilomycin A1, (H) chloroquine, (G) ammonium chloride, or vehicle control: DMSO for 2 hours at 37°C. Cells were then infected with CHIKV 181/25 (MOI=1.0) for 1 hour 37°C, and re-supplemented with media containing compound for a total incubation time of 24 hours. CHIKV infection was evaluated via immunofluorescence by staining for expression of CHIKV capsid protein and total cell count was determined by DAPI staining. Percent infection was determined by determining number of infected cells over total cells. Cell viability was determined via MTT assay. Data for percent (%) infection and cell viability of compounds is represented as normalized to DMSO vehicle control. (I) IC50 and CC50 values and corresponding dose-response curve for viral infectivity and cell viability was determined via non-linear [inhibitor] vs normalized response – variable non-linear slope analysis. Data represented as means ± SEM. Antiviral and cell viability assays were performed in technical triplicate (n=3 donors).

Article Snippet: Other compounds targeting micropinocytosis (EIPA), clathrin (pitstop 2), and dynamin (dynasore) were obtained from MedChemExpress.

Techniques: Cell Culture, Incubation, Control, Infection, Immunofluorescence, Staining, Expressing, Cell Characterization, MTT Assay

Journal: Journal of Virology

Article Title: Bovine coronavirus enters HRT-18 cells via membrane fusion and clathrin-mediated endocytosis in a low pH-, dynamin-, cholesterol-, microtubule-, Rab7-, and Rab11-dependent manner

doi: 10.1128/jvi.01274-25

Figure Lengend Snippet: BCoV entry into HRT-18 cells depends on dynamin. ( A ) The maximum safe concentrations of dynasore were determined using the CCK-8 assay. ( B ) Western blot analysis was used to evaluate the BCoV N protein expression levels, with grayscale analysis performed and presented as a bar graph. ( C ) RT-qPCR was performed to assess the BCoV gene copy numbers. ( D ) TCID 50 assay was used to measure the BCoV viral titers in the cell supernatant. ( E ) IFA was used to detect the number of BCoV-infected cells. Scale bar = 100 µm. ( F ) RT-qPCR was used to evaluate the effect of dynasore on the viral entry. ( G ) RT-qPCR was used to evaluate the effect of dynasore on the viral attachment. ( H ) The siRNA silencing efficiency of dynamin was screened; the effects of dynamin-silenced cells on BCoV infection were assessed by ( I ) Western blot, ( J ) RT-qPCR, ( K ) TCID 50 , and ( L ) IFA. ( M ) RT-qPCR was used to evaluate the effect of dynamin-silenced cells on the viral entry; ( N ) RT-qPCR was used to evaluate the effects of dynamin-targeting siRNA on BCoV attachment. Data are presented as the mean ± SD of three independent experiments (not significant, P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: The inhibitors used in this study included SSAA09E3 (Cat HY-138102, MedChemExpress), a novel inhibitor that blocks the fusion of the viral membrane with the host cell membrane; CPZ (Cat C0982, Sigma), a clathrin-mediated endocytosis inhibitor; nystatin (Cat 475914, Sigma), a caveolae inhibitor that acts as a sterol-binding agent disrupting caveolae; blebbistatin (Cat 203391, Sigma), an inhibitor of micropinocytosis; dynasore (Cat T1848, TargetMol), a dynamin inhibitor; MβCD (Cat T4072, TargetMol), a cholesterol depletion inhibitor; chloroquine (Cat S6999, Selleck) and NH 4 Cl (Cat A9434, Sigma), a potent inhibitor of V-ATPase and a specific inhibitor of acidification of endosomal vesicles; colchicine (Cat HY-16569, MedChemExpress), which inhibits the polymerization of tubulin; E64d (Cat S7393, Selleck), a cathepsin inhibitor; and camostat (Cat HY-13512, MedChemExpress), a TMPRSS2 inhibitor.

Techniques: CCK-8 Assay, Western Blot, Expressing, Quantitative RT-PCR, Infection

Journal: Communications Biology

Article Title: Proteolysis targeting chimera (PROTAC)-driven antibody internalization of oncogenic cell surface receptors

doi: 10.1038/s42003-024-07439-0

Figure Lengend Snippet: Antibody internalization was evaluated in the presence of endocytosis inhibitors (Dyngo-4a and Pitstop2) and ubiquitin-proteasome system inhibitors (TAK-243, PYZD-4409, and MG132). a BT-474 and SKBR3 cells were assayed by in vitro live-cell imaging for Tmab internalization and PROTAC SJF1528 in presence of the dynamin mediated endocytosis inhibitor Dyngo-4a at (30 and 15 µM). b SKBR3 and HCC827 GR6 cells were also assayed for Tmab or anti-MET internalization and PROTAC SJF1528 or 48-284, respectively, in presence of the Uba1 inhibitors TAK-243 and PYZD-4409. c Time course evaluation of the internalized Tmab by Western blots with antibodies specific against light and heavy human IgG chains and HER2 expression on SKBR3 cells treated with or without SJF1528 200 nM and Tmab 2 µg/mL, and SJF1528 200 nM with Tmab 2 µg/mL and MG132 5 µM. d Representative images of immunofluorescence staining of BT-474 cells assayed overnight with SJF1528 200 nM and labeled Tmab 2 µg/mL or SJF1528 200 nM with Tmab 2 µg/mL, MG132 5 µM and a secondary antibody for human IgG–FITC conjugated. Data shown here are representative experiments, every condition has been done in triplicate and lines and error bars represent the medians and SEM. Individual data points are available in the file and the uncropped Western blots are in the Supplementary figures. Statistical significance was evaluated with GraphPad Prism 10 by unpaired t -test and two-tailed p value. **** P < 0.0001.

Article Snippet: Both, the dynamin I/II or clathrin inhibitors Dyngo-4a and Pitstop2 (Med Chem Express HY-13863 and HY-115604, respectively) and Uba1 inhibitors TAK-243 and PYZD-4409 (Med Chem Express HY-100487 and HY-13297, respectively) were added to the cells at the same time that labeled antibodies Tmab and anti-MET were added, and internalization evaluated by acquiring images with IncuCyte every 30 min for 24–48 h as before.

Techniques: In Vitro, Live Cell Imaging, Western Blot, Expressing, Immunofluorescence, Staining, Labeling, Two Tailed Test

Journal: Medical microbiology and immunology

Article Title: The pathogenic responses elicited during exposure of human intestinal cell line with Giardia duodenalis excretory-secretory products and the potential attributed endocytosis mechanism

doi: 10.1007/s00430-024-00806-y

Figure Lengend Snippet: Unless otherwise specified, Caco-2 cells were exposed to Giardia ESPs for the indicated time periods. (A) Upon exposure of IECs with the amount of ESPs of 20, 40 and 100 μg/mL, the mRNA levels of dynamin, clathrin and caveolin-1 were assessed by qPCR analysis. The comparisons were made with the first group of the graph. (B-D) Cells were exposed to ESPs at the concentration of 20 μg/mL. (B) The expression levels of clathrin and caveolin-1 were determined by western blot analysis. (C) Anti-ESP PAB was used to indicate the bonding between ESPs and IECs after a 3-h exposure (scale bar = 20 μm). (D) Binding between ESPs and clathrin/caveolin-1 was assessed by co-IP analysis. All experiments were repeated four times. Data from triplicate wells from a representative of four independent experiments are presented as means ± SD. * p < 0.05, ** p < 0.01.

Article Snippet: We used dynamin inhibitor dynasore (100 μM in use; Abmole, Houston, USA), clathrin inhibitor chlorpromazine (CPZ) (10 μM; Abmole, Houston, USA), and caveolin-1 inhibitor genistein (100 μM; Selleck, Shanghai, China) in inhibition analyses.

Techniques: Activation Assay, Concentration Assay, Expressing, Western Blot, Binding Assay, Co-Immunoprecipitation Assay

Journal: Medical microbiology and immunology

Article Title: The pathogenic responses elicited during exposure of human intestinal cell line with Giardia duodenalis excretory-secretory products and the potential attributed endocytosis mechanism

doi: 10.1007/s00430-024-00806-y

Figure Lengend Snippet: Dynasore was dissolved in 0.1% DMSO for use and shown as “dynasore + DMSO” panel here. Caco-2 cells were exposed to Giardia ESPs at the concentration of 20 μg/mL for 3 h. (A) Dynamin inhibition by dynasore blocked the interactions between ESPs and IECs as assessed by immunofluorescence analysis (scale bar = 20 μm). (B and C) Dynamin inhibition attenuated ESPs-induced cleavage of CASP-3, CASP-9 and PARP and elevation of Bax to Bcl-2 ratio as assessed by western blot and gray value analyses. (C) “Control” was compared to “ESPs”, and “ESPs” was compared to “ESPs + dynasore + DMSO”. (D) Dynamin inhibition suppressed ESPs-induced IEC apoptosis as assessed by AO/EB staining (scale bar = 100 μm). (E) Dynamin inhibition suppressed ESPs-induced upregulation of intracellular NLRP3 and TNF-α and released IL-1β and IL-18 as assessed by western blotting. (F) Dynamin inhibition protected claudin-1, claudin-4, occludin and ZO-1 expressions from being influenced by ESP exposure. (G and H) Dynamin inhibition blocked ESPs-induced elevation of LC3-II to LC3-I ratio and reversed ESPs-induced downregulation of p62 as assessed by western blot and gray value analyses. (H) “Control” was compared to “ESPs”, and “ESPs” was compared to “ESPs + dynasore + DMSO”. All experiments were repeated three times. Data from triplicate wells from a representative of three independent experiments are presented as means ± SD. * p < 0.05, ** p < 0.01.

Article Snippet: We used dynamin inhibitor dynasore (100 μM in use; Abmole, Houston, USA), clathrin inhibitor chlorpromazine (CPZ) (10 μM; Abmole, Houston, USA), and caveolin-1 inhibitor genistein (100 μM; Selleck, Shanghai, China) in inhibition analyses.

Techniques: Concentration Assay, Inhibition, Immunofluorescence, Western Blot, Control, Staining