Figure 1
From: Salt inducible kinases 2 and 3 are required for thymic T cell development
SIK levels in T cell subsets. (A) Naïve CD4 and CD8 T cells were isolated by FACS and subject to TMT labelled proteomic analysis as described in33. For ex vivo stimulations of the TCR, GP33 (glycoprotein amino acids 33–41), 20 ng/ml IL-2 and 2 ng/ml IL-12 were used to stimulate CD8 cells from P14 mice. OVA peptide loaded onto antigen presenting cells was used to stimulate CD4 cells from OT-II mice as described in33. Graphs show mean and standard deviation for SIK2 and SIK3. SIK1 was not detected. For CD4 cells, data are from independent preparations from 3 mice per genotype and for CD8 cells from 6 mice per genotype. Data is derived from Howden et al. 2019, Nat. Immunol. 20, 1542–155433. (B) Thy1+ve thymocytes pooled from 5 mice were sorted based on CD4 and CD8 expression into CD8 single positive, CD4 single positive, double negative (DN) and CD4/8 double positive (DP) cells by FACS. 5 μg of lysate was separated by SDS-PAGE and blotted using the indicated antibodies. Molecular weight markers are shown, with weights in kDa.
