rab7 d95f2  (Cell Signaling Technology Inc)

Journal: PLOS Pathogens

Article Title: Differential roles of the type I and II secretion systems for the intracellular ABC141 Acinetobacter baumannii infection, which elicits an atypical hypoxia response in endothelial cells

doi: 10.1371/journal.ppat.1013265

Figure Lengend Snippet: (A) Representative confocal immunofluorescence microscopy images of Rab5, EEA1, Rab7, CD63 and LAMP1 (red) recruited or not to the ABC141 (yellow) ACVs in EA.hy926 endothelial cells at 10, 20, 30 min or 1, 2, 6 and 24h. DAPI was used to visualize nuclei (blue) and bacteria. (B) Quantification of the percentage of ACVs positive for V-ATPase at different time points after infection. Data correspond to the means ± SD of 3 independent experiments. (C) Representative images of V-ATPase positive (red) ABC141 (white) ACVs at 30 min (left) and 24h post-infection (right). DAPI was used to visualize nuclei and bacteria. All scale bars correspond to 5 μm.

Article Snippet: , Rab7 (D95F2) , Rabbit , 1:200 , Cell signaling 9367T.

Techniques: Immunofluorescence, Microscopy, Bacteria, Infection

Journal: Microbiology Spectrum

Article Title: Cholesterol-rich lipid rafts mediate endocytosis as a common pathway for respiratory syncytial virus entry into different host cells

doi: 10.1128/spectrum.01192-25

Figure Lengend Snippet: RSV entry into cells triggers actin polymerization and endosome formation by cholesterol-rich lipid rafts. ( A ) RSV enters cells by actin-mediated endocytosis. HEp-2 or A549 cells were infected with RSV A2 (MOI = 30) (90 min, 37°C) and fixed (4% PFA, 15 min, room temperature). Cell nuclei were labeled with DAPI (cyan), lipid rafts with Alexa 647-CTB (blue), cholesterol with NBD-cholesterol (green), and F-actin with CellMask orange actin tracking dye (yellow), and images were acquired using confocal microscopy. Scale bar = 10 µm. Intensity profiles show colocalization along the direction of the yellow line. Pearson’s correlation between lipid rafts, cholesterol or F-actin, and RSV. n = 16 cells per group. Error bars represent a 95% confidence interval. ( B ) RSV trafficking from early to late endosomes. HEp-2 or A549 cells were infected with RSV A2 (MOI = 30) (150 min, 37°C) and fixed. Cell nuclei were labeled with DAPI (cyan), lipid rafts with Alexa 647-CTB (blue), and cholesterol with NBD-cholesterol (green). Immunodetection of RSV F (blue), RSV-FITC (green), EEA1, Rab7, and STX 6 (red) was performed using confocal microscopy. Scale bar = 10 µm. Intensity profiles show colocalization along the direction of the yellow line. Pearson’s correlation between lipid rafts, cholesterol or F-actin, and RSV. n = 16 cells per group. Error bars represent a 95% confidence interval.

Article Snippet: The Flotillin-1 (D2V7J) rabbit mAb (#18,934T), caveolin-1 (D46G3) rabbit mAb (#3267), EEA1 (C45B10) rabbit mAb (#3288), Rab7 (D95F2) rabbit mAb (#9367), Rab7 (E9O7E) mouse mAb (#95746), and syntaxin 6 (C34B2) rabbit mAb (#2869) were purchased from Cell Signaling Technology.

Techniques: Infection, Labeling, Confocal Microscopy, Immunodetection

Journal: Journal of Virology

Article Title: IFITM proteins are key entry factors for porcine epidemic diarrhea coronavirus

doi: 10.1128/jvi.02028-24

Figure Lengend Snippet: Interaction and colocalization of the PEDV S protein with IFITM proteins. ( A ) Co-IP assays were performed to assess the interaction between the PEDV S1 protein and human or porcine IFITM proteins. HEK293T cells were cotransfected with plasmids encoding PEDV S1-Fc and various HA-tagged IFITM proteins. At 24 h post-transfection, the cells were harvested. Immunoprecipitation was conducted using an Fc tag antibody, and the presence of IFITM proteins was detected by Western blotting (IB: Anti-HA). Whole-cell lysates (WCLs) were analyzed to confirm expression levels (IB: Anti-Fc, Anti-HA, Anti-β-Tubulin). The Co-IP was repeated three times, yielding similar results. ( B ) Confocal microscopy images showing the intracellular localization of porcine IFITM1 in LLC-PK1 cells are presented. IFITM1 (red) colocalizes with clathrin (green), EEA1 (green), Rab7 (green), and LAMP7 (green), as indicated in the merged images. DAPI (blue) was used to stain the nuclei. ( C ) Confocal microscopy images demonstrating the colocalization of the PEDV S protein (red) with HA-IFITM1 (green) in LLC-PK1 cells are shown. The merged image shows the nuclei stained with DAPI (blue).

Article Snippet: Additional antibodies included EEA1 mouse monoclonal antibody (cat#: 68065-1-Ig), HA tag mouse monoclonal antibody (cat#: 66006-2-Ig), Rab7 (D95F2) XP rabbit monoclonal antibody (cat#: 9367s), and CLTC monoclonal antibody (cat#: 66487-1-Ig) from Proteintech.

Techniques: Co-Immunoprecipitation Assay, Transfection, Immunoprecipitation, Western Blot, Expressing, Confocal Microscopy, Staining