Fig. 3: PRMT5 interacts with snail and the NuRD complex.

A Immunoaffinity purification of PRMT5-containing protein complexes. Cellular extracts from HeLa cells stably expressing HA (control) or HA-PRMT5 were immunopurified with anti-HA affinity columns and eluted with HA peptide. The eluates were resolved by SDS-PAGE and silver stained. The protein bands were retrieved and analyzed by mass spectrometry. Detailed results of the mass spectrometric analysis are provided in Supplementary File 1 (Supplemental Table S4). B Western blotting analysis of purified fractions using antibodies against the indicated proteins. C–E Association of PRMT5 with Snail and the NuRD complex. Whole-cell lysates from HeLa, Ca ski, and SiHa cells were prepared and co-IP was performed with antibodies against the indicated proteins. Immunocomplexes were then tested by IB using antibodies against the indicated proteins. IgG served as a negative control. F Association of Snail with the NuRD complex. Whole-cell lysates from HeLa, Ca ski, and SiHa cells were prepared and co-IP was performed with antibodies against the indicated proteins. Immunocomplexes were then tested by IB using antibodies against the indicated proteins. G Equal amounts of HeLa cellular extracts were immunoprecipitated with antibodies against MTA1 or MTA3 followed by IB with antibodies against the indicated proteins. H Cofractionation of Snail, PRMT5, and the NuRD complex by FPLC. Nuclear extracts of HeLa cells were fractionated on a DEAE Sepharose column followed by a Superose 6 gel filtration column. The fractions were analyzed by western blotting. Molecular weight standards are shown on top (in kDa).