ma104  (Procell Inc)

Journal: bioRxiv

Article Title: Ursolic Acid Inhibits Rotavirus Replication Through Modulation Of Lipid Droplet Homeostasis

doi: 10.64898/2026.04.13.718265

Figure Lengend Snippet: (A) MA104 cells were pre-treated with DMSO or 10 μM UA for 1 h and then infected with different RV strains SA-11, RRV, and NCDV at an MOI of 1 in the presence of the compounds. At 15 h post infection (p.i.), the supernatants were collected, and infectious particles were titrated using the plaque assay method. Each boxplot represents the mean titer after DMSO or 10 μM UA treatment of three independent experiments. The means were analyzed by a Student’s t-test (*p< 0.05, **p < 0.01). (B) MA104 cells were pre-treated with DMSO or 10 μM UA for 1 h, and then the LD accumulation was induced with 100 µM OA for 2 h. Then, the cells were incubated with Bodipy, fixed, and analyzed by CLSM. The left panel shows representative microscopy images, and the boxplots represent the mean number and size of LD of three independent experiments. The means were analyzed by a Student’s t-test (**p < 0.01). The scale bar represents 10 μm. (C-F) Physical characterization of lenses upon UA incorporation, obtained from compression isotherms and imaging of EPC:TG:UA monolayers (C) Lateral pressure at which TG are segregated from the film (π EX ) determined at the sharp kink on the isotherms. (D) Lateral radius ( i.e , from top view of the monolayer) of each individual lens. (E) Lens density (number of lenses/μm 2 ). ( F) Thickness of each individual lens, computed from BAM reflectivity using the equation included in Materials and Methods section. Statistical significance was evaluated for each %UA against the control of %UA=0 using the Tukey test (*p<0.05, **p<0.01, and ***p<0.001). C-F plots were generated using SigmaPlot for Windows v15 (Systat, USA).

Article Snippet: MA104 Clone 1 cells (ATCC CRL-2378.1), derived from Cercopithecus aethiops monkey kidney epithelium, were kindly provided by Dr. Poncet (I2BC, Paris, France).

Techniques: Infection, Plaque Assay, Incubation, Microscopy, Imaging, Control, Generated

Journal: bioRxiv

Article Title: Ursolic Acid Inhibits Rotavirus Replication Through Modulation Of Lipid Droplet Homeostasis

doi: 10.64898/2026.04.13.718265

Figure Lengend Snippet: (A) MA104 cells were treated with 100 µM OA for 2 h and then treated with DMSO or 10 µM UA for 1 h. Then, the cells were incubated with Bodipy, fixed, and analyzed by CLSM. The left panel shows representative microscopy images. The scale bar represents 10 µm. The boxplots represent the mean number and size of LD of three independent experiments. The means were analyzed by a Student’s t-test (**p < 0.01, ****p<0.0001). (B) MA104 cells were treated with 100 µM OA for 2 h and then treated with DMSO or 10 µM UA during 1 h as in (A). Additionally, cells were treated with DMSO or 10 µM UA alone. Then, they were incubated with Bodipy, and the fluorescence intensity was analyzed by flow cytometry. 10,000 events per condition were evaluated from four independent experiments. The left panel shows representative histograms, where the area under the curve corresponds to the fluorescence intensity. The boxplot represents the fluorescence intensity means for each condition. The means were analyzed by a Student’s t-test (***p<0.001; ****p< 0.0001). (C) MA104 cells were treated with 100 µM OA for 2 h and then incubated with DMSO or 10 µM UA during 0, 2, 4, or 6 h. Then, the cells were incubated with Bodipy, fixed, and analyzed by CLSM. The upper panel shows representative microscopy images where the scale bar represents 10 µm. The boxplots represent the mean number and size of LD of three independent experiments. The means were analyzed by a paired-wise Student’s t-test (**p < 0.01, ***p < 0.001).

Article Snippet: MA104 Clone 1 cells (ATCC CRL-2378.1), derived from Cercopithecus aethiops monkey kidney epithelium, were kindly provided by Dr. Poncet (I2BC, Paris, France).

Techniques: Incubation, Microscopy, Fluorescence, Flow Cytometry

Journal: bioRxiv

Article Title: Ursolic Acid Inhibits Rotavirus Replication Through Modulation Of Lipid Droplet Homeostasis

doi: 10.64898/2026.04.13.718265

Figure Lengend Snippet: MA104 EGFP-NSP5 cells were infected with RV at an MOI of 1 and analyzed at 0, 2, 4, and 6 h post infection (p.i.). Then, the cells were incubated with the LD red, fluorescent probe Lipid Tox, fixed, and analyzed by CLSM. The VP were evidenced by the green, EGFP-derived fluorescent dots. The left panel shows representative microscopy images where the scale bar represents 10 µm. The boxplots on the right side represent the mean number and size of LD and VP of three independent experiments. The means were analyzed by a paired-wise Student’s t-test (***p < 0.001, ****p < 0.0001).

Article Snippet: MA104 Clone 1 cells (ATCC CRL-2378.1), derived from Cercopithecus aethiops monkey kidney epithelium, were kindly provided by Dr. Poncet (I2BC, Paris, France).

Techniques: Infection, Incubation, Derivative Assay, Microscopy

Journal: bioRxiv

Article Title: Ursolic Acid Inhibits Rotavirus Replication Through Modulation Of Lipid Droplet Homeostasis

doi: 10.64898/2026.04.13.718265

Figure Lengend Snippet: MA104 cells were treated with DMSO or 10 μM UA for 1 h, then infected with RV at an MOI of 1 and left until 0, 2, 4, and 6 h post infection (p.i.) in the presence of DMSO (left panel) or 10 μM UA (right panel). We colored the LD in green with Bodipy and used anti-NSP2 antibodies to detect the VP. The scale bar represents 10 µm. The boxplots below represent the mean number and size of LD and VP in three independent experiments. The means were analyzed by a paired-wise Student’s t-test (*p < 0.1, **p < 0.01, ***p < 0.001).

Article Snippet: MA104 Clone 1 cells (ATCC CRL-2378.1), derived from Cercopithecus aethiops monkey kidney epithelium, were kindly provided by Dr. Poncet (I2BC, Paris, France).

Techniques: Infection

Journal: bioRxiv

Article Title: Ursolic Acid Inhibits Rotavirus Replication Through Modulation Of Lipid Droplet Homeostasis

doi: 10.64898/2026.04.13.718265

Figure Lengend Snippet: (A) MA104 cells were treated with 10 μM UA for 1, 2, 4, 6, or 8 h. Cells were then processed for Western blot analysis, and GAPDH protein levels were detected. Ponceau S staining was used as a loading control. The boxplot was generated from four independent experiments. Statistical significance was assessed by one-way ANOVA followed by Tukey’s multiple comparisons test (**p < 0.01, ***p < 0.001, ****p < 0.0001). (B) MA104 cells were pre-treated with DMSO or 10 μM UA and infected with RV at an MOI of 1 in the presence of the compounds. At 0, 2, 4, and 6 h post-infection (p.i.), cells were collected and processed for Western blot analysis. OA was used as a control for lipid overload and lipolytic stimulation, whereas the combination of TOFA and OA was used as a control for induction associated with inhibition of lipid synthesis. To confirm cellular infection, RV VP8 protein levels were detected. Ponceau S staining was used as a loading control. The boxplots shown were generated from four independent experiments. Statistical significance was assessed using Student’s t -test (*p < 0.1, **p < 0.01). (C) MA104 cells were pre-treated with DMSO or 10 μM UA and infected with RV at an MOI of 1 in the presence of the compounds. Infection was stopped at 0, 2, 4, and 6 h post-infection (p.i.), and cells were incubated with Bodipy, fixed, and immunostained with an anti-PLIN2 antibody followed by an Alexa Fluor 555-conjugated secondary antibody. Images were acquired by CLSM. The figure shows representative images from three independent experiments. Scale bar represents 10 µm.

Article Snippet: MA104 Clone 1 cells (ATCC CRL-2378.1), derived from Cercopithecus aethiops monkey kidney epithelium, were kindly provided by Dr. Poncet (I2BC, Paris, France).

Techniques: Western Blot, Staining, Control, Generated, Infection, Inhibition, Incubation

Journal: bioRxiv

Article Title: Ursolic Acid Inhibits Rotavirus Replication Through Modulation Of Lipid Droplet Homeostasis

doi: 10.64898/2026.04.13.718265

Figure Lengend Snippet: (A) First row: MA104 cells were left untreated (Control), incubated for 2 h with starvation medium (Stv.), or starvation medium plus Bafilomycin A1 (Stv. + BafA1). Second row: MA104 cells were DMSO- or 10 µM UA-treated for 2 h, in the absence or presence of Bafilomycin A1 (UA + BafA1). Cells were fixed and processed for immunofluorescence staining of LC3 (red), and nuclei were counterstained with Hoechst (blue). Representative CLSM images are shown. The number of LC3-positive puncta (LC3+ dots) per cell was quantified and is shown on the right as boxplots. Data were obtained from four independent experiments. Statistical significance was assessed using Student’s t -test (*P < 0.05; **P < 0.01). (B) Controls (first three lines): MA104 cells were left untreated (Control), incubated for 2 h with starvation medium (Stv.), or starvation medium plus Bafilomycin A1 (Stv. +BafA1) to verify the autophagy modulation. Mock-treated or infected with RV strain SA-11 (MOI = 1) in the presence of DMSO or 10 µM UA. At the indicated times post-infection (0, 2, 4, and 6 h p.i.), cells were collected and processed for Western blot analysis to detect LC3-I and LC3-II. Tubulin was used as a loading control. Representative immunoblots are shown (left). Densitometric quantification of LC3-II normalized to tubulin is shown on the right as boxplots. Data correspond to four independent experiments. Statistical significance was assessed using Student’s t-test (*p < 0.05; **p < 0.01; ***p < 0.001).

Article Snippet: MA104 Clone 1 cells (ATCC CRL-2378.1), derived from Cercopithecus aethiops monkey kidney epithelium, were kindly provided by Dr. Poncet (I2BC, Paris, France).

Techniques: Control, Incubation, Immunofluorescence, Staining, Infection, Western Blot

Journal: bioRxiv

Article Title: Species-specific regulation of porcine STING stability and antiviral signaling via its K61 mediated K48 ubiquitination and proteasome degradation

doi: 10.64898/2026.03.26.714395

Figure Lengend Snippet: (A-B) HEK293T cells were co-transfected with mCherry-pSTING (20 ng) or its mutants together with pcGAS-HA (20 ng), plus ISRE-luc (10 ng) or NF-κΒ-luc (10 ng) and pRL-TK plasmid (0.4 ng) for 24 h. Luciferase activities were detected, and protein expressions were analyzed. (C-F) 3D4/21 cells in 24-well cell crawl slides were co-transfected with Flag-pSTING or K61R mutant together with KDEL-Red (C), mApple-SiT (D), MITO-GFP (E) or Lamp1-GFP (F) (each 0.5 μg) for 24 h, and then cells were fixed, stained and the colocalization of STING and its K61R mutant with organelles was examined by confocal fluorescence microscopy. (G) HEK293T cells in 6-well were transfected with Flag-pSTING or K61R mutant (1 μg/mL) for 24h, and stimulated with 2’3’-cGAMP (2 μg/mL) for 2 h. Then interaction between STING and TBK1/IRF3 was identified by Co-IP using anti-FLAG antibody, whereas rabbit IgG served as the control. (H-I) STING -/- 3D4/21 cells in 12-well were transfected with pmCherry-C1, mCherry-pSTING or K61R mutant (1 μg) for 24 h, and stimulated with 2’3’-cGAMP (2 μg/mL) for indicated times (H) or 6 h (I). Cells were harvested and analyzed by Western blotting (H) and RT-qPCR (I). (J-M) STING -/- 3D4/21 cells were transfected with Flag-pSTING or its mutants (1μg), and then stimulated with 2’3’-cGAMP (2 μg/mL) for 2 h, followed by infections with HSV-1 (MOI=0.01, 36 h) or PRV (MOI=0.01, 24 h). The viral titers in the supernatants from HSV-1 (J) or PRV (L) infected cells was measured by plaque assay, and the HSV-1 and PRV gD gene expressions were measured by RT-qPCR (K and M). ( N-P ) STING -/- MA104 cells were transfected and stimulated as in STING -/- 3D4/21 cells, followed by infections ASFV-GFP, infected cells were detected by fluorescence microscopy and cytometry, Western blotting and quantitative PCR. * p < 0.05 and ** p < 0.01.

Article Snippet: HEK293T cells (ATCC cat # CRL-3216), Vero cells (ATCC cat # CCL-81), Raw264.7 cells (ATCC cat # TIB-71), MDBK cells (ATCC cat # CCL-22) and MA104 cell (ATCC cat # CRL-2378.1) were cultured in DMEM (Hyclone Laboratories, USA) containing 10% fetal bovine serum (FBS, Vazyme Biotech Co., Ltd) and 100 IU/ml of penicillin plus 100 μg/ml streptomycin.

Techniques: Transfection, Plasmid Preparation, Luciferase, Mutagenesis, Staining, Fluorescence, Microscopy, Co-Immunoprecipitation Assay, Control, Western Blot, Quantitative RT-PCR, Infection, Plaque Assay, Cytometry, Real-time Polymerase Chain Reaction