Supplementary Figure 2: Confirmation of the knockdown of AMPKα and TAB1 by lentiviral vector in senescent CD27–CD28– CD4+ T cells. | Nature Immunology

Supplementary Figure 2: Confirmation of the knockdown of AMPKα and TAB1 by lentiviral vector in senescent CD27–CD28– CD4+ T cells.

From: The kinase p38 activated by the metabolic regulator AMPK and scaffold TAB1 drives the senescence of human T cells

Supplementary Figure 2

(a) Extended map and (b) schematic representation of the p-Siren HIV lentiviral vector system used for gene knock-down. The shRNA of interest was cloned downstream the U6-promoter between the BamHI and EcoRI restriction sites. Measurement of (c) AMPKα and (d) TAB1 expression by both immunoblot (top) and quantitative PCR (bottom) in highly differentiated CD27- CD28- CD4+ T cells transduced with either shAMPKα or shTAB1, as indicated. An irrelevant scrambled short hairpin (shCtrl) expressing vector was used as internal control. Immunoblots are representative of 3 independent experiments. Samples for quantitative PCR were analyzed from 4 different donors and normalized against housekeeping GAPDH. Knockdown efficiency was evaluated 96 hours post-transduction (day 7). **p<0.01 and ***p< 0.001 values were assessed by a paired Student’s t test. Errors bars depict s.e.m.

Back to article page