Fig. 5

SMN, PLS3, hnRNP F, and hnRNP H1/2 associate in a large complex. a SMN and PLS3 independently associated with hnRNP H1/2 based on co-immunoprecipitation from HEK293T cells transfected with tagged proteins. b Endogenous PLS3, hnRNP F, and hnRNP H 1/2 associated with SMN in mouse brain extracts, based on immunoprecipitation with antisera against endogenous SMN protein. Arrowheads indicate the band of interest in blots with multiple bands. Heavy and light chain bands are labeled when present. c Endogenous SMN, hnRNP F, and hnRNP H1/2 associated with endogenous PLS3 using the same approach outlined in b. Arrowheads indicate the band of interest in blots with multiple bands. Heavy and light chain bands are labeled when present. IP on endogenous mouse brain extract was done from 3 independent brain extractions and preparations. d Size fractionation studies of whole cell lysates from HEK293T cells showed that actin, SMN, PLS3, and hnRNP F and/or H1/2 proteins co-fractionate in complexes of the indicated size (bracket), although the bulk of hnRNP F/H1/2 protein is in other smaller complexes. HEK293T cells were transfected with PLS3-Flag, and hnRNP F-GFP, endogenous hnRNP H1/2, actin, and SMN were detected using antisera