(a) Solubilized membrane extracts from CLV1-GFP co-expressed with BAM2-FLAG in transient expression were IPd with anti-GFP antibodies and co-IPs were detected with anti-FLAG antibodies. A dilution series of 10%, 5% and 1% of total bound and unbound fractions were assayed on protein gel blots to estimate the efficiency of IP and co-IP (see Experimental Procedures).
(b) CRN-GFP co-expressed with CLV2-MYC in transient expression was IPd with anti-GFP antibodies and co-IPs were detected with anti-MYC antibodies. A dilution series of 2%, 1% and 0.2% of total bound and unbound fractions were assayed on protein gel blots to estimate the efficiency of IP and co-IP.
(c) CLV1-FLAG co-expressed with CLV2-MYC in transient expression was IPd with anti-FLAG antibodies and co-IPs were detected with anti-MYC antibodies. A dilution series of 8%, 4%, 1%, 0.4% and 0.1% of total bound, and a dilution series of 4%, 1%, 0.4%, 0.1%, and 0.04% of unbound fractions were assayed on protein gel blots to estimate the efficiency of IP and co-IP.
(d) CLV1-FLAG co-expressed with CLV2-MYC and CRN-GFP in transient expression was IPd with anti-FLAG antibodies and co-IPs were detected with anti-MYC. A dilution series of 8%, 4%, 1% and 0.4% of total bound, and a dilution series of 4%, 1%, 0.4% and 0.1% of unbound fractions were assayed on protein gel blots to estimate the efficiency of IP and co-IP.