Effects of Recombinant Cytokines and Growth Factors on C/EBPβ Expression, Protease Sensitivity and Subnuclear Localization in Two Macrophage Cell Lines
| Factor (concentration) . | P388D1(IL1) Cells . | IC-21 Cells . | ||
|---|---|---|---|---|
| Resistance to Proteolysis* . | IIF Staining† . | C/EBPβ Induction‡ . | IIF Staining† . | |
| Conditioned medium | + | D | + | D |
| IL-1α (10 ng/mL) | − | P | + | P |
| IL-1β (10 ng/mL) | − | P | + | P |
| IL-6 (20 ng/mL) | − | P | + | P |
| TNF-α (10 ng/mL) | ND | ND | + | P |
| IL-1ra (20 ng/mL) | − | P | − | − |
| IL-2 (10 ng/mL) | − | P | − | − |
| IL-3 (10 ng/mL) | − | P | − | − |
| IL-4 (10 ng/mL) | − | P | − | − |
| IL-7 (10 ng/mL) | − | P | − | − |
| IL-8 (25 ng/mL) | − | P | − | − |
| IL-10 (20 ng/mL) | − | P | − | − |
| IL-12 (50 U/mL) | − | P | − | − |
| Oncostatin M (20 ng/mL) | − | P | − | − |
| LIF (10 ng/mL) | − | P | − | − |
| IFN-α (500 U/mL) | − | P | − | − |
| IFN-γ (500 U/mL) | − | P | − | − |
| M-CSF (10 ng/mL) | − | P | − | − |
| G-CSF (10 ng/mL) | − | P | − | − |
| GM-CSF (5 ng/mL) | − | P | − | − |
| HGF (5 U/mL) | − | P | − | − |
| bFGF (10 ng/mL) | − | P | − | − |
| TGF-β (10 ng/mL) | − | P | − | − |
| LPS (20 μg/mL) | − | P | + | ND |
| Factor (concentration) . | P388D1(IL1) Cells . | IC-21 Cells . | ||
|---|---|---|---|---|
| Resistance to Proteolysis* . | IIF Staining† . | C/EBPβ Induction‡ . | IIF Staining† . | |
| Conditioned medium | + | D | + | D |
| IL-1α (10 ng/mL) | − | P | + | P |
| IL-1β (10 ng/mL) | − | P | + | P |
| IL-6 (20 ng/mL) | − | P | + | P |
| TNF-α (10 ng/mL) | ND | ND | + | P |
| IL-1ra (20 ng/mL) | − | P | − | − |
| IL-2 (10 ng/mL) | − | P | − | − |
| IL-3 (10 ng/mL) | − | P | − | − |
| IL-4 (10 ng/mL) | − | P | − | − |
| IL-7 (10 ng/mL) | − | P | − | − |
| IL-8 (25 ng/mL) | − | P | − | − |
| IL-10 (20 ng/mL) | − | P | − | − |
| IL-12 (50 U/mL) | − | P | − | − |
| Oncostatin M (20 ng/mL) | − | P | − | − |
| LIF (10 ng/mL) | − | P | − | − |
| IFN-α (500 U/mL) | − | P | − | − |
| IFN-γ (500 U/mL) | − | P | − | − |
| M-CSF (10 ng/mL) | − | P | − | − |
| G-CSF (10 ng/mL) | − | P | − | − |
| GM-CSF (5 ng/mL) | − | P | − | − |
| HGF (5 U/mL) | − | P | − | − |
| bFGF (10 ng/mL) | − | P | − | − |
| TGF-β (10 ng/mL) | − | P | − | − |
| LPS (20 μg/mL) | − | P | + | ND |
Cells were grown for 16 to 20 hours in fresh medium containing the indicated dose of each factor and then subjected to C/EBPβ analysis.
Abbreviations: LIF, leukemia inhibitory factor; IFN, interferon; M-CSF, macrophage-colony stimulating factor; GM-CSF, granulocyte/macrophage-colony stimulating factor; HGF, hepatocyte growth factor; bFGF, basic fibroblast growth factor; TGF-β, transforming growth factor-β; ND, no data.
C/EBPβ resistance to proteolysis in P388D1(IL1) cells was assayed by EMSA as described in Fig 1 (−, p14-C/EBPβ EMSA complex; +, p34-C/EBPβ complexes).
IIF (indirect immunofluorescence) staining of C/EBPβ was performed as described in Fig 3 (D, diffuse staining; P, punctate staining; −, weak or undetectable staining).
C/EBPβ induction refers to increased levels of C/EBPβ protein in IC-21 nuclear extracts as determined by Western blotting (+, elevated levels of C/EBPβ; −, basal levels of C/EBPβ).