rab7 d95f2 rabbit mab  (Cell Signaling Technology Inc)

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: Cells

Article Title: AMPK Activation Downregulates TXNIP, Rab5, and Rab7 Within Minutes, Thereby Inhibiting the Endocytosis-Mediated Entry of Human Pathogenic Viruses.

doi: 10.3390/cells14050334

Figure Lengend Snippet: Figure 7. AMPK activation downregulates Rab5, Rab7, and TXNIP rapidly. Confocal microscopy of Rab5 (magenta), Rab7 (magenta), and TXNIP (magenta) after SA (A,C), AMPK (A), ULK activator (A), and cycloheximide (B) treatment. (C) Analyses of the time-dependent downregulation of Rab5, Rab7, and TXNIP. Scale bars: 20 µm.

Article Snippet: Antibodies: Anti-PVR (CD155) Antibody, clone 4B3, ZooMab® Rabbit Monoclonal recombinant, Cells 2025, 14, 334 4 of 22 Sigma Aldrich; anti-LDL Receptor Antibody, clone 2N19, ZooMab® Rabbit Monoclonal recombinant, Sigma Aldrich (Taufkirchen, Germany); anti-Rab5A Antibody #2143, Cell Signaling; anti-Rab7 (D95F2) XP® Rabbit mAb #9367, Cell Signaling; and anti-TXNIP Antibody, clone 1K14 ZooMAb® Rabbit Monoclonal, Sigma Aldrich; anti-β-Actin Antibody, clone 6L12, ZooMAb® Rabbit Monoclonal, Sigma Aldrich; anti-SARS-CoV-2 Spike Protein (S1-NTD) Antibody #56996, Cell Signaling, anti-SARS-CoV/SARS-CoV-2 NSP8 Monoclonal Antibody (5A10), Invitrogen; and anti-LAMP1 (D4O1S) Mouse mAb #15665, Cell Signaling; anti-GRP78 Polyclonal Antibody, Invitrogen.

Techniques: Activation Assay, Confocal Microscopy

Journal: bioRxiv

Article Title: A Novel TRPC6 Mutation Causes Autosomal Dominant FSGS

doi: 10.1101/2025.02.11.637765

Figure Lengend Snippet: iPS-cell derived podocytes have reduced expression of slit diaphragm proteins, ER congestion, and reduced lysosomes. Panel A shows representative immunoblots of podocyte markers including Nephrin, Synaptopodin, TRPC6, Podocin, CD2AP, GLEPP1, and WT1, the housekeeping protein GAPDH. Panel B shows the quantification of immunoblots for the podocyte markers. Data represents three biologically independent experiments and is defined as ± standard error of the mean. We performed one-way ANOVA with Dunnett’s multiple comparison post hoc analyses with a family-wise alpha threshold of 0.05. Panel C shows electron micrograph of differentiated podocytes with protein aggresomes in the ER of Duke 5 and Duke 7 podocytes. Panel D shows LAMP1 vesicles, Podocin, and TRPC6 expression in podocytes. The LAMP1 vesicle number remains unchanged in the podocytes. Panel E shows lysosomal biogenesis-associated RAB7 expression in the podocytes. RAB7 expression was reduced in the mutants. Data is representative of three biologically independent experiments and the data is defined as ± standard error of the mean. We performed one-way ANOVA with Sidak’s multiple comparison post hoc analyses with a family-wise alpha threshold of 0.05. Scale bar: Panel C, top row 1000 µm, bottom row 500 nm; Panel D, top row 20 µm, bottom row 5 µm; Panel E, 100 µm.

Article Snippet: The primary antibodies used included guinea pig Anti-Nephrin (ARP; #GPN02, 1:500), rabbit Anti-Podocin (Abcam, mAb #ab50339, 1:1000), rabbit Anti-TRPC6 (Abcam, pAb #ab228771) and mouse Anti-CD2AP (B-4) (Santa Cruz, #sc-25272), Calnexin (C5C9) Anti-Rabbit (Cell Signaling, mAb #2679), LAMP1 (D2D11) XP Anti-Rabbit (Cell Signaling, mAb #9091), RAB7 (D95F2) XP and Anti-Rabbit mAb (Cell Signaling, mAb #9367).

Techniques: Derivative Assay, Expressing, Western Blot, Comparison

Journal: Current Biology

Article Title: Systems mapping of bidirectional endosomal transport through the crowded cell

doi: 10.1016/j.cub.2024.08.026

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-Rab7 (D95F2) , Cell signaling Technology , Cat#9367; RRID: AB_1904103.

Techniques: Recombinant, CRISPR, Sequencing, Introduce, Cloning, Software