Extended Data Fig. 1: Generation of Lck variant mouse models.
From: Unique roles of co-receptor-bound LCK in helper and cytotoxic T cells
(a) A scheme of targeting of the Lck locus for the generation of the LckCA allele by CRISPR/Cas9. Resulting LckCA (repaired via homologous recombination according to the DNA template) and LckKO (non-homologous end joining causing a short deletion) alleles and the respective LCK protein sequences are shown. (b) A scheme of targeting of the Lck locus for the generation of the LckKR allele by CRISPR/Cas9. Resulting LckKR (repaired via homologous recombination according to the DNA template) allele and the respective LCK protein sequence are shown. (c) LCK protein levels in thymi and LNs of Lck variant mice were detected by immunoblotting showing the comparable expression of Lck in these strains. β-actin staining was used as a loading control. LAT staining was used as a control for T-cell derived proteins in the lysates. A representative experiment out of 3 in total is shown. (d-e) Immunoprecipitation of CD4 (d) and CD8β (e) followed by immunoblotting and LCK staining in indicated Lck variant mice. A representative experiment out of 2 (d) or 3 (e) in total is shown.
