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. 2021 Aug 13;17(8):e1009753. doi: 10.1371/journal.pgen.1009753

Fig 2. MTL5 interacts with most components of the MuvB core complex.

Fig 2

(A) Silver staining of the samples immunoprecipitated with MTL5 antibody (epitope from 221 aa to 475 aa) from pachytene and diplotene spermatocytes of 10-week-old wild-type mouse after SDS-PAGE. Rabbit IgG served as a negative control. Red stars mark the different bands between IgG and MTL5 antibody group. Blue arrowheads indicate the unspecific bands. M indicates protein marker. (B) Identification of proteins interacting with MTL5 by IP-MS followed by subtractive analysis of three proteomic data sets with two biological repetitions. Proteins with at least three unique peptides in wild-type groups, but absent in both IgG and knockout (KO) groups, were considered as candidate MTL5-interactive proteins. Numbers in the brackets indicate the number of proteins. (C) Rank of the top candidates, from high to low, sorted by the coverage (%). The top 6 candidates are shown. (D) Western blot detection of the interaction between MTL5 (fused with a MYC tag) and each of MuvB core complex components or MYBL1 (fused with a GFP tag) in HEK293T cells after co-immunoprecipitation with MYC-tag magnetic beads. pEGFP-N1 was served as the negative control. IP, immunoprecipitation. IB, immunoblotting.