Skip to main content
. 2019 Jun 19;47(15):7886–7900. doi: 10.1093/nar/gkz526

Figure 2.

Figure 2.

Characterization of the Arabidopsis ILS complex. (A) Size exclusion chromatography analysis of soluble proteins extracted from inflorescence tissues of plants simultaneously expressing pILP1::ILP1-GFP, p35S::NTR1-10xMYC and p35S::PRP43b-3xFLAG. Proteins were analyzed by Western blot, whereas U1, U5 and U6 RNAs were checked by Northern blot analysis. (B) Reciprocal bimolecular fluorescence complementation (BiFC) analyses among ILP1, NTR1 and PRP43b. Scale bar = 20 μm. Paired constructs were transiently expressed in Arabidopsis leaf protoplasts, and fluorescence was measured 24 h after transfection. Percentage means positive BiFC ratio for corresponding protein pairs. Values (n) indicate the number of counted protoplasts. See also Supplementary Figure S2A for results at lower magnification. (C–E) Reciprocal co-immunoprecipitation (co-IP) assays among ILP1-GFP, NTR1-10xMYC and PRP43b-3xFLAG using ILP1-GFP (C), NTR1-10xMYC (D) and PRP43b-3xFLAG (E) as a bait. Endogenous PRP8 was analyzed using a commercial anti-PRP8 antibody. Asterisk indicates unknown bands. Input = 25% for IPs and 1% for co-IPs, except for the interactions between ILP1-GFP and NTR1-10xMYC, where input was 5% for co-IPs.