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: The American Journal of Pathology

Article Title: The D2.B10- Dmd mdx /J Mouse Model of Duchenne Muscular Dystrophy Exhibits a Severe Mitochondrial Deficiency Not Observed in the C57BL/10ScSn- Dmd mdx /J Mouse

doi: 10.1016/j.ajpath.2025.09.005

Figure Lengend Snippet: Relative protein expression changes to the latent transforming growth factor (TGF)-β–binding protein 4 (LTBP4) pathway and annexin A6 (ANXA6). LTBP4 is a known genetic modifier in human patients with Duchenne muscular dystrophy and impacts TGF-β signaling. TGF-β acts on SMAD4, which directly interacts with mitochondrial cytochrome c oxidase subunit 2 (MTCO2) and can cause mitochondrial fragmentation via dynamin-related protein 1 (DRP1). A – L: All proteins were assessed for expression levels ( A – F ) relative to total protein ( G – L ). H: TGF-β levels were elevated in D2-WT mice compared with B10-WT mice, and complex IV subunit 2 (MTCO2) was elevated in the D2- mdx mice compared with D2-WT. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Antibodies against latent TGF-β–binding protein 4 (LTBP4; Novus Biologicals, Centennial, CO; NBP2-43671; 1:500), TGF-β (Novus Biologicals; NBP2-46108; 1:2000), annexin A6 (ANXA6; ProteinTech, Rosemont, IL; 68086-1-Ig; 1:2000), mitochondrial cytochrome c oxidase subunit 2 (MTCO2; Thermo Fisher; A-6404; 1:500), SMAD4 (ProteinTech; 10231-1-AP; 1:100), dynamin-related protein 1 (DRP1; ProteinTech; 12957-1-AP; 1:1000), and Total OXPHOS Rodent WB Antibody Cocktail (Abcam; ab110413; 1:25,000) were used.

Techniques: Expressing, Binding Assay

Journal: bioRxiv

Article Title: Ca 2+ and DRP1 drive endocytic lysosome reformation at tripartite contact sites

doi: 10.64898/2026.01.30.702748

Figure Lengend Snippet: (A) Immunoblot for the phosphorylation of mTOR effector proteins during different conditions. Torin was used as a positive control for mTOR inhibition. Tubulin was used as a loading control. (B) Quantification of the fold change p-S6K/S6K from immunoblot data in (A). Data represents the mean of n=4 replicates. one-way ANOVA using Tukey’s multiple comparison, **P= 0.0011 (Untreated VS Torin), ns P = 0.9809 (Untreated VS YM201636), ns P = 0.99997 (Untreated VS 15 min recovery), ns P = 0.6835 (Untreated VS 30 min recovery), ns P = 0.99997 (Untreated VS 60 min recovery). (C) Quantification of the fold change of p-S6K/S6K for from immunoblot data in (A). Data represents the mean of n=4 replicates. one-way ANOVA using Tukey’s multiple comparison, ***P = 0.000038 (Untreated VS Torin), ns P = 0.9370 (Untreated VS YM201636), ns P = 0.99999 (Untreated VS 15 min recovery), ns P = 0.999993 (Untreated VS 30 min recovery), ns P = 0.9605 (Untreated VS 60 min recovery). (D) Live-cell imaging of cells expressing LAMP1-GFP (green) stained with LysoTracker deep red (magenta) to assess the lysosomal reformation in presence of 250 nM torin. Images were acquired after 2 h YM201636 treatment and 30 min washout. (G) Average area of LAMP1-positive structures was quantified. Each dot represents the average area of per 100 µm 2 ROI; plot shows mean area from a total of 90 ROI from 30 cells across n=3 biological replicates; Unpaired t-test; ns, P = 0.1980 (DMSO vs Torin). (F) Live-cell imaging of cells expressing LAMP1-mScarlet (magenta) together with DNM2-GFP or the GTPase mutant DNM2 (K44A)-GFP to assess DNM2 function in tubule fission. Images were acquired after 2 h YM201636 treatment and 30 min. (G) Quantification of LAMP1-positive tubule length (magenta) in cells overexpressing wild-type or mutant DNM2. Each dot represents the length of a single tubule; plot shows mean length from a total of 150 tubules across n=3 biological replicates; Unpaired t-test; ns P = 0.0564 (DNM2 WT vs. DNM2 K44A). (H) Schematic representation of the proximity labelling assay using LAMP1 fused to the biotin ligase TurboID. (I) Volcano plot comparing the biotinylated proxisome of LAMP1-TiD in cells treated with YM201636 for 2 h plus 30 min recovery versus untreated controls. Proteins significantly enriched during tubulation and fission are highlighted on the right side of the plot. Data from n=3 biological replicates. (J) Major categories of candidate proteins enriched during tubulation and fission.

Article Snippet: The following commercially available plasmids were obtained: LAMP1-GFP (Cat. 34831/ Addgene), LAMP1-mScarlet (Cat. 98827/ Addgene), pSpCas9(BB)–2A-GFP (pX458) (Cat. 48138/ Addgene), pSpCAS9 (BB) 2A-puro (pX459) (Cat. 48139/ Addgene), EGFR-GFP (Cat. 32751/ Addgene), WT Dynamin 2-GFP (Cat. 34686/ Addgene), GFP-Dynamin2 K44A (Cat. 22301/ Addgene), BFP-KDEL (Cat. 49150/ Addgene), mCh-Climp63 (Cat. 136293/ Addgene), pmCherry C1 MFF (Cat. 157760/ Addgene), pCAG-mito-RCaMP1h (Cat. 105013/ Addgene).

Techniques: Western Blot, Phospho-proteomics, Positive Control, Inhibition, Control, Comparison, Live Cell Imaging, Expressing, Staining, Mutagenesis