Anti dlst

Total cell lysates from CD4⁺ T cells, were obtained through incubation of cells for 20 min at 4 °C in a solution of 50 mM Tris-HCl (pH 7.5), 150 mM NaCl and 1.0% Triton X-100, plus SigmaFast protease inhibitor (S8820; Sigma-Aldrich) and Sigma phosphatase inhibitor (P5726; Sigma-Aldrich), and immunoblot analyses were performed using the following antibodies: anti-aldolase, anti-enolase 1, anti-hexokinase I, anti-DLAT, anti-DLST (all 1:1000 dilution and from Cell Signaling Technology, Beverly, MA) anti-Glut-1 (1:500 dilution and from Abcam) and anti- VDAC (1:1000 dilution and from Santa Cruz Biotechnology). The filters were also probed with an ERK1/2 antibody (1:1000 dilution from Santa Cruz Biotechnology) to normalize for the amount of loaded protein.

Total cell lysates and Western blot analysis were performed as previously described (De Rosa et al., 2007 (link)). The antibodies used were the following: anti-aldolase, anti-enolase, anti-hexokinase, anti-PKM1/2, anti-FAS, anti-apoA4, anti-HADHA, anti-ACAD9, anti-CPT1A, anti-DLST, anti-SDHA, anti-phospho STAT5, anti-STAT5, anti-phospho-S6, anti-S6 (all 1:1000 dilution and from Cell Signaling Technology), anti-FoxP3 (1:500 dilution from eBioscience). anti-ERK1/2 (1:500 dilution) and anti-phospho-ERK1/2 (1:500 dilution) (from Santa Cruz Biotechnology), The filters were also probed with an actin antibody (1:1000 from Santa Cruz) to normalize for the amount of loaded protein. All filters were quantified as previously described (De Rosa et al., 2007 (link)).