Journal: BMC Immunology

Article Title: TGFβ signaling plays a critical role in promoting alternative macrophage activation

doi: 10.1186/1471-2172-13-31

Figure Lengend Snippet: Expression of M2 markers on BMDMs is impaired in the absence of TβRII signaling. (A ) Western blot analysis of WT and TβRII -/- BMDMs cultured for 24 hrs with medium alone (M0), with LPS/IFNγ (M1), IL-4 (M2), or hTGFβ1. β-actin serves as a loading control. Increased Arg-1 in KO BMDM upon hTGFβ1stimulation due to residual WT cells, see Additional file : Figure S2E. ( B ) Real-time RT-PCR for the indicated genes after incubation for 24 hrs with medium alone (M0) or IL-4 (M2). ( C ) Real-time RT-PCR for the indicated genes in naïve peritoneal macrophages. Fold change is with respect to the expression level of M0 WT. Solid bar, WT ; Open bar, TβRII -/- peritoneal macrophages. The results shown are representative of one of two independently derived sets of BMDMs from different mice.

Article Snippet: After clotting of blood, serum was harvested by centrifugation at 800 g for 10 min. Serum IFNγ levels were measured using a mouse IFNγ ELISA kit (Biolegend) according to the manufacturer’s protocol.

Techniques: Expressing, Western Blot, Cell Culture, Control, Quantitative RT-PCR, Incubation, Derivative Assay

Journal: BMC Immunology

Article Title: TGFβ signaling plays a critical role in promoting alternative macrophage activation

doi: 10.1186/1471-2172-13-31

Figure Lengend Snippet: Lack of TβRII signaling is correlated with decreased Galectin-3, β-catenin and phospho-Akt levels. ( A ) Real-time RT-PCR for lgals3 in BMDMs. Fold change is with respect to the expression of M0 WT. Solid bar, WT ; Open bar, TβRII -/- . ( B ) Cell surface expression of Galectin-3 on WT and TβRII -/- BMDMs measured using FACS. MFI, mean fluorescent intensity. The result shown is representative of one of two independently derived sets of BMDMs from different mice for both A and B. ( C ) Western blot analysis of β-catenin expression. BMDM were treated with medium alone (M0), LPS/IFNγ (M1), or IL-4 (M2). Results are representative of 3 pairs of matched WT and TβRII -/- mice. Western blot analysis of phosphorylated-Stat6 ( D ) and phosphorylated-Akt ( E ) in WT and TβRII -/- BMDMs that had been treated with IL-4 for 1 hr. Results are shown for 2 independent mice per genotype. The numbers at the bottom represent the band intensity ratio of phosphorylated protein versus total protein, normalized to lane 1 on each blot.

Article Snippet: After clotting of blood, serum was harvested by centrifugation at 800 g for 10 min. Serum IFNγ levels were measured using a mouse IFNγ ELISA kit (Biolegend) according to the manufacturer’s protocol.

Techniques: Quantitative RT-PCR, Expressing, Derivative Assay, Western Blot