Journal: Discover Oncology

Article Title: Integrated WGCNA retrieval of T-cell exhaustion genes related to radiosensitivity in locally advanced cervical cancer and prediction of immunotherapy efficacy

doi: 10.1007/s12672-025-03673-y

Figure Lengend Snippet: IHC assay results of the protein expression levels of APIBEC3H, CCR7, PILRA, and SLC15A3 in LACC patients

Article Snippet: Subsequently, the sections were incubated at 4 °C overnight with the corresponding primary antibody against APOBEC3H (1:200, PA5-54426, Invitrogen, USA), CCR7 (1:100, 25898-1-AP, Proteintech, China), PLRA (1:100, PA5-147474, Invitrogen, USA), SLC15A3 (1:100, PA5-61614, Invitrogen, USA).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Internalization of fluorescein isothiocyanate-conjugated monoclonal anti-human CCR7 induced by recombinant CCL19 in intact HEK 293a cells that express recombinant CCR7. nt, non-transfected cells.

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Recombinant, Transfection

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Endocytosis of the anti-myc monoclonal antibody (clone 4A6, conjugated to AlexaFluor-488, final concentration in the culture medium 3.3 nM) as determined by co-treatment with the CCL19-myc construction in intact HEK 293a cells that optionally and transiently expressed CCR7. A control conditioned medium (CM) or authentic CCL19 were used as control stimuli. The undiluted CM were transferred for a 30-min incubation period before rinsing and observation. Original magnification ×1000.

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Concentration Assay, Control, Incubation

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Cytofluorometry of HEK 293a cells that optionally expressed CCR7, sequentially detached and further stimulated with the undiluted CM of other cells producing CCL19-myc, authentic CCL19, or CM of untransfected cells, as indicated (30 min incubation at 37°C). Left: distributions based on the counting of 10,000 cells. A threshold of autofluorescence was defined using control cells with no fluorophore. It was surpassed only under one set of experimental conditions (arrow). Right: mean fluorescence of cells in replicated experiment. ANOVA indicated that the values were heterogeneous ( p < 10 -4 ). * p < 0.01 vs. top-most value (control CM, untransfected recipient cells) by Dunnett’s test.

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Incubation, Control, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Epifluorescence microscopy studies in cells co-expressing β-arrestin 1 -Cherry and CCR7 and stimulated as indicated (stimulant and duration). Stimulation always included the fluorescent anti-myc monoclonal antibody 4A6 (green signal). Either CCL19-myc CM or authentic CCL19 rarely condensed β-arrestin 1 at the level of plasma membranes of endosomes (arrowhead indicates possible co-localization with the CCL19-myc-antibody cargo). Cells that expressed well the red-emitting transgene were assumed to co-express co-transfected CCR7 and were evaluated for morphology ( n = 16–49 eligible cells per group). The average number of red condensed structures was 0.4 ± 0.4 in cells treated with control CM and did not vary significantly in all other groups (Kruskal–Wallis test). The average number of green condensed structures in cells treated with control CM was 0 per cell and varied significantly according to treatments ( p < 10 -4 , Kruskal–Wallis test; p values for Dunn’s multiple comparison test for each value vs. that of control CM reported thereafter). There were 8.3 ± 1.7 green specs per cell ( p < 0.05), 25.8 ± 3.2 ( p < 0.001), 33.1 ± 4.4 ( p < 0.001), and 0 (N.S.) in cells treated with CCL19-myc CM for 5, 10, or 30 min, or with recombinant CCL19, respectively.

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Epifluorescence Microscopy, Expressing, Clinical Proteomics, Transfection, Control, Comparison, Recombinant

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Epifluorescence microscopy studies in cells co-expressing CCR7 and β-arrestin 2 -GFP and stimulated as indicated (stimulant and duration). Stimulation did not include the fluorescent anti-myc antibody. Either CCL19-myc CM or authentic CCL19 frequently condensed β-arrestin 2 at the level of plasma membrane or endosomes. Right: number of green intracellular condensed structures (“specs”) per cell ± SEM. Numbers close to bars indicate the number of evaluated HEK 293a cells. The Kruskal–Wallis test indicated that the values were heterogenous ( p < 10 -4 ). The effect of each treatment vs. the effect of control CM was evaluated using Dunn’s multiple comparison test. * p < 0.01; ** p < 0.001.

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Epifluorescence Microscopy, Expressing, Clinical Proteomics, Membrane, Control, Comparison

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Epifluorescence microscopy studies in cells co-expressing one of three forms of Rab5-Cherry fusion proteins and CCR7 and stimulated for 30 min with either CCL19-myc or control CM. Co-localization between Rab5-positive corpuscles and the CCL19-myc-antibody cargo was observed (arrowheads). The dominant positive (GTP-locked) mutant of the fusion protein produced typical giant vacuoles that included the green light-emitting cargo. The dominant negative Rab5-GDP-locked-Cherry inhibited the endocytosis of the green cargo (see text for numerical analysis).

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Epifluorescence Microscopy, Expressing, Control, Mutagenesis, Produced, Dominant Negative Mutation

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Epifluorescence microscopy studies in cells co-expressing Rab7-Cherry and CCR7 and stimulated as indicated (stimulant and duration). Co-localization between Rab7-positive corpuscles and the CCL19-myc-antibody cargo was not highly frequent (arrowheads), but increased in frequency as a function of incubation duration (see text for numerical analysis).

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Epifluorescence Microscopy, Expressing, Incubation

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Presence of CCR7 in A375 melanoma cells. (A) RT-PCR for CCR7 in the human melanoma A375 cell line. (B) c-Fos induction in A375 cells stimulated as indicated for 1 h (typical immunoblot and histograms representing the densitometry of replicated experiments).

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Pharmacology

Article Title: C-C chemokine receptor-7 mediated endocytosis of antibody cargoes into intact cells

doi: 10.3389/fphar.2013.00122

Figure Lengend Snippet: Detection of endogenous CCR7 in A375 cells using detection of the endocytosed CCL19-myc-4A6 antibody complex using theTyramide Signal Amplification (TSA) system that enzymatically generates AlexaFluor-488 labeling. Epifluorescence and transmission, original magnification: ×1000. Right: proportion of cells with a mean fluorescence intensity above a set threshold (Photoshop level ≥ 50/255) in large photographic records of A375 cells stained as in the left of figure (the number of evaluated cells is indicated next to each histogram). The proportions were compared with that of control cells without antibody (top-most histogram, χ 2 test; * p < 10 -4 ).

Article Snippet: Using these cells, a first CCR7 imaging strategy was based on staining intact and live HEK 293a cells previously transfected with the vector encoding for the receptor with carboxyfluorescein-conjugated monoclonal anti-human CCR7 (CD197; clone 150503, final concentration 5 μg/ml; R&D Systems, Minneapolis, MN, USA).

Techniques: Amplification, Labeling, Transmission Assay, Fluorescence, Staining, Control

Journal: Developmental cell

Article Title: The Vascular Niche Regulates Hematopoietic Stem and Progenitor Cell Lodgment and Expansion via klf6a-ccl25b.

doi: 10.1016/j.devcel.2017.07.012

Figure Lengend Snippet: Figure 7. The Enhancement of Ccl21/Ccr7 Signaling on HSPC Expansion Is Conserved in Mice (A) The expression of Klf6, Ccl21 and Ccr7 in sorted ECs (CD31+ CD45 TER119) and LSKs (LinSca-1+c-Kit+) from E14.5 mouse fetal liver by qPCR (mean ± SEM, t test; **p < 0.01, ***p < 0.001, ns [not significant] > 0.05, n = 3). (B) Immunofluorescence analysis of mouse Ccr7 expression in E14.5 fetal liver. The white arrow- heads indicate Ccr7 and Runx1 double-positive cells. Scale bar, 25 mm. (C) The flow chart of ex vivo HSPC culture. (D) CFU-C assay of HSPCs co-cultured with Ccl21 at 10 ng/mL and 40 ng/mL compared with control, respectively (mean ± SEM, t test; *p < 0.05, **p < 0.01, ***p < 0.001, ns > 0.05, n = 3). (E) CFU-C assay of HSPCs cultured in MyeloCult M5300 medium containing Ccl21 at 40 ng/mL and the neutralizing anti-Ccl21 antibody or neutral- izing anti-Ccr7 antibody to block the Ccl21/Ccr7 signaling, respectively (mean ± SEM, t test; ***p < 0.001, n = 3). (F) Model for development of HSPCs in the CHT niche. Klf6a+ vascular ECs can produce ccl25b that attracts HSPCs to settle closely to the caudal vein, and perivascular ECs are triggered by an arriving HSPC to form an EC pocket. Later, HSPC proliferate in the stem cell pocket in the CHT ni- che. The orange cells indicate perivascular ECs and green cells indicate HSPCs. The red plots mark chemokine ligand (ccl25b).

Article Snippet: For antibody blocking experiments, 500 HSPCs were cultured in MyeloCultTMM5300medium, containing Ccl21 at 40 ng/ml and the neutralizing anti-Ccl21 antibody (R&D systems, AF457) at 2.5 ug/ml or neutralizing anti-Ccr7 antibody (R&D systems, MAB3477) at 5 ug/ml, to block the endogenous Ccl21/Ccr7 signaling, respectively.

Techniques: Expressing, Ex Vivo, Cell Culture, Control, Blocking Assay