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. 2009 Jun 30;7(6):e1000139. doi: 10.1371/journal.pbio.1000139

Figure 7. RIN4 is expressed in guard cells.

Figure 7

Guard cell protoplasts (GCPs) were purified from Col 0 leaves and visually inspected for purity by light microscopy. Half of the GCP sample was used for RNA extraction and half for total protein extraction. (A) RNA was isolated from entire Arabidopsis leaves or GCPs and subjected to RT-PCR. RIN4 mRNA is highly expressed in guard cells. The expression of phosphoenolpyruvate carboxylase 2 (ATPPC2, At2g42600), which has low-level expression in guard cells and high-level expression in mesophyll cells served a control for guard cell protoplast purity [47]. Actin (AtACT2) served as a loading control. (B) Anti-RIN4 immunoblots detected RIN4 protein expression in both Col 0 leaf tissue and GCPs. Thirty µg of total protein extract was loaded per lane. (C) RNA samples from (A) were subjected to RT-PCR to detect the expression of additional innate immune signaling components. EDS1, PAD4, RPS2, NDR1, EFR, and CERK1 transcript levels were detected in GCPs and leaf tissue after a 28-cycle amplification. +, RT-PCR, −, no-RT control.