Journal: PLoS ONE

Article Title: Evidence of the Presence of a Functional Dot/Icm Type IV-B Secretion System in the Fish Bacterial Pathogen Piscirickettsia salmonis

doi: 10.1371/journal.pone.0054934

Figure Lengend Snippet: The immunofluorescence was made 5 days post-infection. Lysosomes were stained in red with LysoTracker Red reagent and P. salmonis was detected with a FITC conjugated antibody. A: CHSE-214 cell line infected with P. salmonis . B: Sf21 cell line infected with P. salmonis . C: RTS11 cell line infected with P. salmonis . D: CHSE-214 cell line infected with formaldehyde-inactivated P. salmonis , the immunofluorescence stain was made at 48 hours after infection.

Article Snippet: For a highly proliferative intracellular environment, it has been used the Sf21 insect cell line (Invitrogen) since it has been described for producing high titers of P. salmonis at 15 days post-infection .

Techniques: Immunofluorescence, Infection, Staining

Journal: PLoS ONE

Article Title: Human Osteoarthritic Cartilage Shows Reduced In Vivo Expression of IL-4, a Chondroprotective Cytokine that Differentially Modulates IL-1β-Stimulated Production of Chemokines and Matrix-Degrading Enzymes In Vitro

doi: 10.1371/journal.pone.0096925

Figure Lengend Snippet: Primers for PCR analysis.

Article Snippet: The antibodies used were the same as those used for confocal analysis, IL-4Rα (Mouse R&D MAB230), IL-13Rα1 (Goat R&D AF152), IL-2Rγ (Goat R&D AF284) used at 10 µg/ml and detected by Donkey anti-Mouse Dy Light 647 or Donkey anti-Goat Dy Light 647 at 15 µg/ml.

Techniques:

Journal: PLoS ONE

Article Title: Human Osteoarthritic Cartilage Shows Reduced In Vivo Expression of IL-4, a Chondroprotective Cytokine that Differentially Modulates IL-1β-Stimulated Production of Chemokines and Matrix-Degrading Enzymes In Vitro

doi: 10.1371/journal.pone.0096925

Figure Lengend Snippet: A : a representative example of IL-4Rα and IL-2Rγ colocalization (IL-4 Receptor type I) in control and OA cartilage. B : a representative example of IL-4Rα and IL-13Rα1 (IL-4 Receptor type II) in control and OA cartilage (bar = 10 µm). Panels show IL-4R staining (Dy Light 647, rendered in red), either IL-2Rγ or IL-13Rα (Alexa 555, rendered in white), nuclear counterstaining (sybr green, rendered in green) and merged signals.

Article Snippet: The antibodies used were the same as those used for confocal analysis, IL-4Rα (Mouse R&D MAB230), IL-13Rα1 (Goat R&D AF152), IL-2Rγ (Goat R&D AF284) used at 10 µg/ml and detected by Donkey anti-Mouse Dy Light 647 or Donkey anti-Goat Dy Light 647 at 15 µg/ml.

Techniques: Staining, SYBR Green Assay

Journal: PLoS ONE

Article Title: Human Osteoarthritic Cartilage Shows Reduced In Vivo Expression of IL-4, a Chondroprotective Cytokine that Differentially Modulates IL-1β-Stimulated Production of Chemokines and Matrix-Degrading Enzymes In Vitro

doi: 10.1371/journal.pone.0096925

Figure Lengend Snippet: A: SOX-9 protein expression in high density cultures compared to proliferating chondrocytes by western blot analysis. The housekeeping protein GAPDH is used as a loading control. A representative example out of the four examined. B: Cumulative evaluation of IL-4 receptor subunit gene expression in high-density chondrocytes in 5 different patients. Values represent the number of molecules per 100,000 GAPDH as assessed by real time PCR. Boxes indicate the 25% and 75% percentiles, whiskers indicate the minimum to maximum values, and bars indicate the median. C: flow cytometric analysis of the level of expression of each IL-4R subunit in high density culture OA chondrocytes. Dark pattern: isotype control; white pattern: Dy Light 647 signal referring to IL-4Rα (left), IL-2Rγ (middle) and IL-13Rα (right). Each curve refers to 5,000 cells. D representative example of IL-4Rα and IL-2Rγ colocalization (IL-4 Receptor type I) in high-density cultures of OA chondrocytes (bar = 25 µm). On the right, higher magnifications showing IL-4R staining (Dy Light 647, rendered in red in d4), IL-2Rγ (Alexa 555, rendered in white in d3), nuclear counterstaining (sybr green, rendered in green in d2) and merged signals (d1).

Article Snippet: The antibodies used were the same as those used for confocal analysis, IL-4Rα (Mouse R&D MAB230), IL-13Rα1 (Goat R&D AF152), IL-2Rγ (Goat R&D AF284) used at 10 µg/ml and detected by Donkey anti-Mouse Dy Light 647 or Donkey anti-Goat Dy Light 647 at 15 µg/ml.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Staining, SYBR Green Assay