Journal: Innate immunity

Article Title: Identification of CD300a as a new hypoxia-inducible gene and a regulator of CCL20 and VEGF production by human monocytes and macrophages.

doi: 10.1177/1753425913507095

Figure Lengend Snippet: Figure 6. Differential modulation of CCL20 and VEGF production by CD300a cross-linking on primary Mn. Mn were seeded onto plates precoated with a specific anti-CD300a agonist mAb (+) or a control IgG1 (–) and goat anti-mouse IgG-F(ab0)2, and cultured for 24 h under hypoxic conditions (Hypo, +) or in the presence of DFX (+). Cells were harvested and analyzed for (A) CCL20 and (C) VEGF mRNA expression by qRT-PCR. Relative transcript expression was calculated as detailed in the legend to Figure 1B. Data shown are expressed as fold increase relative to control Ig-cross-linked cells (considered equal to 1) and are representative of one of three experiments performed. (B) CCL20 and (D) VEGF content in conditioned medium were assayed by ELISA. Results are expressed as pg/106 cells/ml and represent the mean SEM of three different experiments. Values significantly different from those of cells cross- linked with isotype-matched Ab according to the Student’s t-test: *P 0.05; **P 0.01.

Article Snippet: Flow cytometry was performed as described.38 For the detection of surface markers, cells were re-suspended with FACS buffer (PBS supplemented with 0.2% BSA, 0.01% NaN3) and incubated with the fluorochrome-conjugated mAbs, anti-CD14-PE and antiCD300a (IRp60, clone P192 produced as described in Cantoni et al.14), the isotype-matched control Abs, mouse IgG2a-PE (Biolegend, Campoverde, Milano, Italy) and mouse IgG2a (R&D Systems, Space Import Export, Milano, Italy), and the secondary FITC-conjugated goat anti-mouse Ab (Invitrogen Life Technologies, Monza, Italy) for 30min at 4 C, after blocking nonspecific sites with rabbit IgG (Sigma, Milano, Italy).

Techniques: Control, Cell Culture, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Innate immunity

Article Title: Identification of CD300a as a new hypoxia-inducible gene and a regulator of CCL20 and VEGF production by human monocytes and macrophages.

doi: 10.1177/1753425913507095

Figure Lengend Snippet: Figure 7. CCL20 and VEGF production regulation by CD300a triggering on MDMs. MDMs were incubated with anti-CD300a Ab (+) or control isotype IgG1 (–) and goat anti-mouse IgG-F(ab0)2 for 24 h under hypoxic conditions (Hypo, +) or in the presence of DFX (+). (A) CCL20 mRNA expression, (C) VEGF mRNA expression and (B, D) cytokine release into the supernatant were analyzed 24 h after stimulation, as detailed in the legend to Figure 6. Values significantly different from those of cells cross-linked with isotype- matched Ab according to the Student’s t-test: * P 0.05.

Article Snippet: Flow cytometry was performed as described.38 For the detection of surface markers, cells were re-suspended with FACS buffer (PBS supplemented with 0.2% BSA, 0.01% NaN3) and incubated with the fluorochrome-conjugated mAbs, anti-CD14-PE and antiCD300a (IRp60, clone P192 produced as described in Cantoni et al.14), the isotype-matched control Abs, mouse IgG2a-PE (Biolegend, Campoverde, Milano, Italy) and mouse IgG2a (R&D Systems, Space Import Export, Milano, Italy), and the secondary FITC-conjugated goat anti-mouse Ab (Invitrogen Life Technologies, Monza, Italy) for 30min at 4 C, after blocking nonspecific sites with rabbit IgG (Sigma, Milano, Italy).

Techniques: Incubation, Control, Expressing

Journal: bioRxiv

Article Title: Myosin 10 uses its MyTH4 and FERM domains differentially to support two aspects of spindle pole biology required for mitotic spindle bipolarity

doi: 10.1101/2023.06.15.545002

Figure Lengend Snippet:

Article Snippet: Secondary antibody , Alexa Fluor ® 647 AffiniPure Goat Anti-Rat IgG (H+L) , Jackson Immuno Research , Cat#112-605-167 , Immunofluorescence (1:400).

Techniques: Recombinant, CRISPR, Imaging, Fluorescence, Labeling, Concentration Assay, Western Blot, Transfection, Cloning, Sequencing, Control, Immunofluorescence, Software