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Figure 3 | BMC Plant Biology

Figure 3

From: Interaction between two rice mitogen activated protein kinases and its possible role in plant defense

Figure 3

In planta interaction of OsMPK20-4 with OsMPK3. (A) HA tagged OsMPK20-4 and Myc tagged OsMPK3 proteins transiently transformed individually or together in tobacco leaves were used for co-immunoprecipitation experiment. The protein from pCAMBIA1302 empty vector infiltrated leaves was used as negative control (Mock). First IP (immunoprecipitation) was carried out using anti-HA antibody then immunoblots (IB) of the precipitated proteins was carried out using anti-c-Myc and anti-HA antibodies. IBs were developed using HRP based chemiluminescent substrate. (B) Sub-cellular co-localization studies were conducted using YFP tagged OsMPK3 and CFP tagged OsMPK20-4 proteins. Localization of the indicated tagged proteins in transiently transformed tobacco leaves was observed using YFP and CFP filters of confocal laser scanning microscope (Leica AOBS system). Bars = 20 μm. (C) Fluorescence resonance energy transfer assay of the tagged proteins was conducted post 48 hr of transient transformation in tobacco leaves. Cyan and yellow are pseudo colors representing CYP and YFP respectively. Pictures were taken by confocal microscope (TCS SP5; Leica). Bars = 20 μm. (D) FRET efficiency (%) as observed in PM (plasma membrane) and nucleus (n = 3).

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