mouse monoclonal allophycocyanin (apc)-conjugated antibodies against cd63 (clone mem-259)  (Thermo Fisher)

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 1. Integrative experimental pipeline towards analyzing the utility of MRC-5 cell-derived exosomes as drug delivery systems of naturally derived anticancer compounds. The cell culture supernatant derived from normal lung fibroblast MRC-5 cells was subjected to initial low-speed centrifugation steps (2,500xg for 15 min, 3,500xg for 15 min, 25,000xg for 25 min) to obtain the exosome production medium (EP-medium) free of “contaminating” live/dead cells, cellular debris, apoptotic fragments and microvesicles. Exosomes were isolated from EP-medium following 3 different methodologies: (a) ultracentrifugation-based method using two sequential high-speed centrifugation steps (2x 135,000xg for 2 h), (b) membrane-based affinity binding method via exoEasy Maxi Kit, (c) a modified protocol employing EP-medium initial ultrafiltration via centrifugal filter units and exosome collection via membrane-based affinity binding method (exoEasy Maxi Kit). The exosomes collected via the (a), (b) and (c) exosome isolation approaches were subjected to morphological and physicochemical characterization using DLS technique for diameter and zeta-potential determination, Bicinchoninic acid assay (BCA) for total exosomal protein quantification and cryo-TEM analysis for exosome visu alization. Based on these data the modified protocol (c) was selected for downstream experimental assays. Specifically, the CD9 and CD63 exosomal protein markers were identified via Western Blotting and bead-based Flow Cytometry analysis. The total miRNA and the total proteomic profiling of isolated exosomes were accomplished via high throughput NGS and nano-LC-MS/MS analysis, respectively. Functional bioinformatic analysis and network construction were performed based on the miRNA and proteomic data. The MRC-5 cell-derived exosomes isolated via the method shown in (c) were labeled with DPPE-rhodamine lipid via the simple incubation method and subjected to cellular uptake assays to investigate the exosomes’ “intrinsic targetability” towards lung adenocarcinoma A549 and squamous tongue carcinoma HSC-3 cells using qualitative CLSM and quantitative flow cytometry analysis. Exosomes were also loaded with Curcumin and/or Doxorubicin via the simple incubation method and sonication-based or lyophilization-based approach, respectively, to investigate the drug encapsulation efficiency of exosomes as well as the pharmacological profile of drug-loaded exosomes via CLSM microscopy and cytotoxicity evaluation.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Derivative Assay, Cell Culture, Centrifugation, Isolation, Membrane, Binding Assay, Modification, Zeta Potential Analyzer, Acid Assay, Western Blot, Flow Cytometry, High Throughput Screening Assay, Liquid Chromatography with Mass Spectroscopy, Functional Assay, Labeling, Incubation, Sonication, Lyophilization, Encapsulation, Microscopy

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 2. Comparative physicochemical characterization and morphological evaluation of MRC-5 cell-derived exosomes isolated via ultracentrifugation (a-c), the membrane-based binding affinity method (exoEasy kit, QIAGEN) (d-f), and a modified protocol employing exosomes concentration via ultrafiltration followed by exosomes isolation via the membrane-based binding affinity method (g-j). Size distribution of exosomes isolated via ultracentrifugation (a), the membrane-based affinity method (d), and the modified protocol (g). Cryo-TEM morphological evaluation of the exosomes isolated via ultracentrifugation (b-c), the membrane- based binding affinity method (e-f), and the modified protocol (h-j). Spherical vesicles (pointed by white arrows) with a 30–60 nm diameter represent the iso lated exosomes. The bright white circles surrounding the exosomal lumen are the lipid bilayer of the exosomal vesicles. Exosome aggregates (pointed by red arrows) generated by high-speed centrifugation are also depicted in the ultracentrifugation-based isolation approach.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Derivative Assay, Isolation, Membrane, Binding Assay, Modification, Concentration Assay, Generated, Centrifugation

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 4. CD63 and CD9 exosomal protein markers identification via Western blotting and flow cytometry. (a) Western blot analysis of common exosomal marker CD63 in MRC-5 cell-derived exosomes (MRC-5 EXO) and corresponding whole cell lysate (MRC-5 WCL). An equal amount of protein (10 μg) from exosomes and whole cell lysate, calculated by BCA protein assay kit, were loaded in wells of 10 % SDS-PAGE gel for separation. (b) Validation of the CD9 exosomal protein expression on the surface of the MRC-5 cell-derived exosomes. The histogram represents the negative control (anti-CD63 capture beads) incubated with the anti-CD9 PE antibody without the addition of exosomes, as well as the MRC-5 cell-derived exosomes captured by the beads with subsequent incubation with the anti-CD9 antibody. (c-d). Dot plots depicting the gating strategy for the negative control and the exosome-containing samples. The flow cytometry analysis was conducted via FlowJo software.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Western Blot, Flow Cytometry, Marker, Derivative Assay, Bicinchoninic Acid Protein Assay, SDS Page, Biomarker Discovery, Expressing, Negative Control, Incubation, Software

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 6. Integrative miRNA and target genes interaction network. Visualization of the miRNAs identified in MRC-5 cell-derived exosomes isolated via the modified protocol and their associated target genes based on miRTarBase database. The genes are depicted in green elliptic shape, and the miRNAs are in orange round rectangles. A red elliptic shape surrounds the “hub” genes targeted by multiple exosomal miRNAs. The network was constructed via the Cytoscape App.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Derivative Assay, Isolation, Modification, Construct

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 8. Determination of the loading efficiency of DOX HCL, CUR, and 16:00 Liss Rhodamine lipid in exosomes derived from the MRC-5 cells. EXO-DOX freeze-drying and EXO-DOX sonication loading methods represent freeze-dried exosomes resuspended in DOX-solution and exosomes sonicated in DOX-solution, respectively. In both methods, a fixed proportion (1:1) of MRC-5 cell-derived exosomal protein with DOX was added. EXO-CUR incubation and EXO-LISS RHO PE incubation represent exosomes incubated with the lipophilic drug curcumin and the fluorescent lipid 16:00 Liss Rhodamine lipid, respectively. CUR loading into exosomes was conducted by mixing a fixed proportion (5:1) of MRC-5 cell-derived exosomes (in PBS) with CUR (dissolved in ethanol and diluted with PBS to reach a final ethanol concentration of 1 %). The DPPE-rhodamine lipid was incorporated in exosomes by mixing a fixed proportion (7:1) of MRC-5 cell-derived exosomes (in PBS) with the fluorescent lipid. (b) Drug-loaded exosome size determination. The excess-free drug was removed via ultracentrifugation, and the exosomal pellet was resuspended and analyzed via DLS. SEM values represent the Standard Error of the Mean of 3 replicates. (c) Images of UC polyallomer tubes after the ultracentrifugation step. White arrows indicate the well-shaped colored exosomal pellet generated by the ultracentrifugation. No pellet was observed in the tubes containing an equal amount of compounds without exosomes (negative controls).

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Derivative Assay, Sonication, Incubation, Concentration Assay, Generated

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 9. MRC-5 cells-derived exosomes uptake efficiencies by MRC-5, A549 and HSC-3 cells. (a) Flow cytometry analysis of MRC-5 cells-derived exosomes inter nalization efficiency by MRC-5 cells. The MRC-5 cells were co-cultured with the DPPE-rhodamine lipid-labeled exosomes for 12 hours and analyzed by flow cytometry. (b) Flow cytometry analysis of MRC-5 cells-derived exosomes internalization efficiency by A549 cells. The A549 cells were co-cultured with the DPPE- rhodamine lipid-labeled exosomes for 12 hours and analyzed by flow cytometry. (c) Flow cytometry analysis of MRC-5 cells-derived exosomes internalization ef ficiency by HSC-3 cells. The HSC-3 cells were co-cultured with the DPPE-rhodamine lipid-labeled exosomes for 12 hours and analyzed by flow cytometry. The grey- colored curve represents untreated-control cells; Light purple-colored curve represents cells incubated with rhodamine-stained exosomal samples; Dark purple- colored curve represents the target cells incubated with a specified amount of DPPE-rhodamine lipid (positive control) (d) The internalization efficiency of MRC- 5 cell derived exosomes by the normal lung MRC-5, the cancer lung A549 cells and the human tongue squamous carcinoma HSC-3 cells, as elucidated by flow cytometry analysis. The uptake rates demonstrate that at least 8.85 %, 23 % and 65 % of the MRC-5, A549 and HSC-3 cells respectively have efficiently internalized the exosomes.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Derivative Assay, Flow Cytometry, Cell Culture, Labeling, Control, Incubation, Staining, Positive Control

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 13. DOX-loaded MRC-5 cell-derived exosomes internalization profile by “homologous” normal MRC-5 cells after co-culturing for 12 hours (scale bar= 10 μm). Red fluorescent spots, indicated by white arrows, represent exosomes loaded with the hydrophilic doxorubicin and incubated with MRC-5 cells. DOX at an amount equal to that of exosomal preparations was ultracentrifuged following the same procedure as for DOX loading experiments and added to the recipient cell conditioned medium providing the controls (a-c). The DOX-loaded exosomes were prepared via freeze-drying and co-cultured with the MRC-5 target cells (d-f). The DOX-loaded exosomes were prepared using the sonication method and co-cultured with the MRC-5 target cells (g-i). The nucleus of the target cells was stained with DAPI and displays blue fluorescence. Samples were observed with CLSM. DAPI, 4′,6-diamidino-2-phenylindole; CLSM, confocal laser scanning microscopy.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Derivative Assay, Incubation, Cell Culture, Sonication, Staining, Fluorescence, Confocal Laser Scanning Microscopy

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 14. DOX loaded MRC-5 cells-derived exosomes internalization profile by “heterologous” tumor A549 cells after co-culturing for 12 hours (scale bar= 10 μm). Red fluorescent spots, indicated by white arrows, represent exosomes loaded with the hydrophilic doxorubicin and incubated with MRC-5 cells. DOX at an amount equal to that of exosomal preparations was ultracentrifuged following the same procedure as for DOX loading experiments and added to the recipient cell conditioned medium providing the controls (a-c). The DOX-loaded exosomes were prepared via freeze-drying and co-cultured with the MRC-5 target cells (d-f). The DOX-loaded exosomes were prepared using the sonication method and co-cultured with the MRC-5 target cells (g-i). The nucleus of the target cells was stained with DAPI and displays blue fluorescence. Samples were observed with CLSM. DAPI, 4′,6-diamidino-2-phenylindole; CLSM, confocal laser scanning microscopy.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Derivative Assay, Incubation, Cell Culture, Sonication, Staining, Fluorescence, Confocal Laser Scanning Microscopy

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 15. DOX loaded MRC-5 cells-derived exosomes internalization profile by “heterologous” tumor HSC-3 cells after co-culturing for 12 hours (scale bar= 10 μm). Red fluorescent spots, indicated by white arrows, represent exosomes loaded with the hydrophilic doxorubicin and incubated with MRC-5 cells. DOX at an amount equal to that of exosomal preparations was ultracentrifuged following the same procedure as for DOX loading experiments and added to the recipient cell conditioned medium providing the controls (a-c). The DOX-loaded exosomes were prepared via freeze-drying and co-cultured with the MRC-5 target cells (d-f). The DOX-loaded exosomes were prepared using the sonication method and co-cultured with the MRC-5 target cells (g-i). The nucleus of the target cells was stained with DAPI and displays blue fluorescence. Samples were observed with CLSM. DAPI, 4′,6-diamidino-2-phenylindole; CLSM, confocal laser scanning microscopy.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: Derivative Assay, Incubation, Cell Culture, Sonication, Staining, Fluorescence, Confocal Laser Scanning Microscopy

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 16. In vitro viability of “autologous” lung normal MRC-5 cells (a), “heterologous” lung tumor A549 (b) and tongue tumor HSC-3 cells (c) after incubation for 48 hours with control medium (control), 1μМ free CUR, 1μМ CUR-loaded in exosomes (4.9 μg exosomal protein), 3μМ free CUR and 3μМ CUR-loaded in exosomes (14.75 μg exosomal protein). 96-well plates were used, and the viability of the cells was assessed by performing CCK-8 assays. The graph was created in GraphPad Prism. One-way ANOVA with Tukey’s multiple comparison test was applied. *p<0.05, **p<0.01, ***p<0.005.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: In Vitro, Incubation, Control, CCK-8 Assay, Comparison

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 17. In vitro viability of “autologous” lung normal MRC-5 cells (a) and “heterologous” lung tumor A549 (b) and tongue tumor HSC-3 cells (c) after incubation for 48 hours with control medium (control), 0.3μМ free DOX, 0.3μМ DOX-loaded in exosomes (3.1 μg exosomal protein), 0.7μМ free DOX and 0.7μМ DOX-loaded in exosomes (7.2 μg exosomal protein). 96-well plates were used, and the viability of the cells was assessed by performing CCK-8 assays. The graph was created in GraphPad Prism. One-way ANOVA with Tukey’s multiple comparison test was applied. *p<0.05.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: In Vitro, Incubation, Control, CCK-8 Assay, Comparison

Journal: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences

Article Title: Towards implementing optimized methodologies and in-depth molecular profiling to assess the pharmacological potential of human lung fibroblast exosomes as bio-shuttles for cancer therapeutics.

doi: 10.1016/j.ejps.2025.107155

Figure Lengend Snippet: Fig. 18. In vitro viability of “autologous” lung normal MRC-5 cells (a) and “heterologous” lung tumor A549 (b) and tongue tumor HSC-3 cells (c) after co-incubation for 48 hours with 0.7μМ DOX loaded in exosomes (7.2 μg exosomal protein) and 1μМ CUR loaded in exosomes (4.9 μg exosomal protein). 96-well plates were used and the viability of the cells was assessed by performing CCK-8 assays. The graph was created in GraphPad Prism. One-way ANOVA with Tukey’s multiple com parison test was applied. *p<0.05.

Article Snippet: Then, the immunoblots were blocked in 1 X PBST (0.1 % Tween 20) supplemented with 5 % skim milk and incubated with the primary monoclonal antibody against CD63 exosomal marker (1:5000 dilution, CD63 Monoclonal antibody, 67,605–1-lg, Proteintech Group, Rosemont, USA) overnight at RT.

Techniques: In Vitro, Incubation, CCK-8 Assay

Journal: International Journal of Nanomedicine

Article Title: Intranasal Delivery of BACE1 siRNA and Berberine via Engineered Stem Cell Exosomes for the Treatment of Alzheimer’s Disease

doi: 10.2147/IJN.S506793

Figure Lengend Snippet: Preparation and identification of hUC-MSC engineered exosomes. ( A ) Representative TEM images of MsEVB@R/siRNA (Scale bar: 100 nm). ( B ) Zeta potentials of MsEV, MsEVB/siRNA, and MsEVB@R/siRNA were analyzed by DLS. ( C ) Particle size of MsEVB@R/siRNA analyzed by DLS. ( D ) Protein expression levels of exosome markers CD63, CD9, TSG101, and GAPDH detected by Western blot (n = 3). ( E ) MsEVB@R/siRNA measured by spectrophotometer at 345 nm wavelength (n = 3). ( F ) In vitro drug release from different formulations (n = 3). ( G ) MsEV, MsEVB/siRNA and MsEVB@R/siRNA on Aβ protein expression in SH-SY5Y cells overexpressing APP gene measured by immunofluorescence and quantitative analysis (Scale bar: 50 μm, n = 3) (DAPI: Blue fluorescence; Aβ: Green fluorescence). ( H ) Relative expression of BACE1 mRNA in SH-SY5Y cells overexpressing APP gene after intervention by MsEV, MsEVB/siRNA, and MsEV@BBR/siRNA (n = 3). ( I and J ) Cell viability of SH-SY5Y cells and BV2 cells measured by CCK8 after incubation with MsEV, MsEVB/siRNA and MsEVB@R/siRNA for 24 h (n = 3). Data were presented as mean ± SD. ** p < 0.01.

Article Snippet: CD9 Rabbit Monoclonal Antibody (AG1418, Beyotime, China), CD63 Rabbit Monoclonal Antibody (AF1471, Beyotime, China), TSG101 Mouse Monoclonal Antibody (AG3472), Anti-amyloid 1–42 antibody, anti-IL-6 antibody, anti-TNF-α antibody, anti-NEUN antibody, and anti-Nestin antibody were purchased from China Servicebio Technology CO., LTD. All chemicals used were of analytical or reagent grade.

Techniques: Expressing, Western Blot, Spectrophotometry, In Vitro, Immunofluorescence, Fluorescence, Incubation

Journal: mBio

Article Title: RAB5 is a host dependency factor for the generation of SARS-CoV-2 replication organelles

doi: 10.1128/mbio.03314-24

Figure Lengend Snippet: Early endosomal membranes highly co-localize with SARS-CoV-2 dsRNA. VeroE6 cells were infected with SARS-CoV-2 HK (MOI = 1) in a synchronized manner. Six hours later, cells were fixed, blocked, and stained for the virus dsRNA and the intracellular markers RAB5, CD63, TGN46, GOSR1, CD81, RAB11, ADRP, MitoTracker, and calnexin. Images: representative pictures of three independent experiments. White scale bar: 10 µm. Graph: the Pearson’s correlation coefficient ( R ) value for the co-localization of SARS-CoV-2 dsRNA and the intracellular markers listed above was calculated from eight randomly selected fields. Raw values with their mean and SEM are represented. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: CD63 , Mouse monoclonal (MEM-259) to CD63 (LAMP-3) , 1:100 , ThermoFisher Scientific, MA1-19281.

Techniques: Infection, Staining, Virus