Page 1 of 9 SUPPLEMENTAL DATA Coordinate Regulation of Bcl11b Activity in Thymocytes by the MAPK Pathways and Protein Sumoylation Ling-juan Zhang, Walter K. Vogel, Xiao Liu, Acharawan Topark-Ngarm, Brian L. Arbogast, Claudia S. Maier, Theresa M. Filtz, and Mark Leid Supplemental Figure S1 Bcl11b phosphorylation sites identified by tandem mass spectrometry (spectra) Supplemental Figure S2 Coverage map of Bcl11b post-translational modifications identified by tandem mass spectrometry Supplemental Table 1 QPCR primers used in this study Supplemental Table 2 Bcl11b co-immunoprecipitating NuRD complex proteins indentified in primary mouse thymocytes
9
Embed
SUPPLEMENTAL DATA Coordinate Regulation of Bcl11b Activity ... · SUPPLEMENTAL DATA Coordinate Regulation of Bcl11b Activity in Thymocytes by the MAPK ... 1548.5 y 13 –P 1449.5
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
Page 1 of 9
SUPPLEMENTAL DATA
Coordinate Regulation of Bcl11b Activity in Thymocytes by the MAPK Pathways and Protein Sumoylation
Ling-juan Zhang, Walter K. Vogel, Xiao Liu, Acharawan Topark-Ngarm, Brian L. Arbogast, Claudia S. Maier, Theresa M. Filtz, and Mark Leid
Supplemental Figure S1 Bcl11b phosphorylation sites identified by tandem mass spectrometry (spectra)
Supplemental Figure S2 Coverage map of Bcl11b post-translational modifications identified by tandem mass spectrometry
Supplemental Table 1 QPCR primers used in this study
SUPPLEMENTAL FIGURE S1. Bcl11b phosphorylation sites identified by tandem mass spectrometry. Ion trap tandem mass spectra were recorded by a Thermo LTQ-FTMS Ultra. Exact mass determinations of the parent ion were derived from full-scan spectra recorded in the ICR cell at a resolution of 100,000 (m/z = 400). Ions resulting from the neutral lose of H2O and NH3 are identified by prefixing º- and ª-symbols while the neutral lose of H3PO4 is identified by a –P suffix in the spectral annotations. Panels A, N, P, and Q are singly phosphorylated peptides where the site of phosphorylation is assigned to multiple sites. Diagnostic phosphosite specific tandem masses are noted in the annotations.
SUPPLEMENTAL FIGURE S2. Coverage map of Bcl11b post-translational modifications identified by tandem mass spectrometry. Combined trypsin and cyanogen bromide/trypsin digests of natively expressed mouse thymus Bcl11b sequenced by tandem spectrometry. Phosphorylated residues are shown in red and the underlined pairs indicate sites to which at least a single phosphorylation is ambiguously assigned. The SUMO1 and SUMO2/3 adduction site, lysine 679, is shown in green. Sequences covered by these experiments are shown in uppercase bold characters. Sequence presented is full-length mouse Bcl11b (Swiss-Prot: Q99PV8) but includes coverage derived from the shortened splice variant missing residues 142–213, isoform 2 (Swiss-Prot: Q99PV8-2).
Page 9 of 9
SUPPLEMENTAL TABLE 1 Primers used for RT-qPCR amplifications
SUPPLEMENTAL TABLE 2 Bcl11b co-immunoprecipitating NuRD complex proteins identified in primary mouse thymocytes NuRD complex proteins in Bcl11b immune complex were identified by mass spectrometry. Primary mouse thymo-cytes were stimulated with PMA and A23187 for the indicated times and immunoprecipitated with anti-Bcl11b antibody as described the Experimental Procedures. Immune complexes were prepared for mass spectrometry analysis after SDS-PAGE size-fractionation (10 fractions/condition). Each in-gel tryptic digest was separately analyzed on an LTQ-FTMS Ultra mass spectrometer and the resulting data sets combined for protein identity analysis; see Experimental Procedures. NuRD complex members identified are shown with their percent sequence coverage and the number of peptides unique to the indicated protein identification shown in parentheses for each experimental determination.