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. 2021 Feb 7;22(4):1673. doi: 10.3390/ijms22041673

Figure 2.

Figure 2

Identification of OsCRP1 target cpRNAs. The GFP-tagged transgenic rice plants were generated using the OsCc1::OsCRP1-GFP vector. Soluble protein and total RNA extractions and RNA-immunoprecipitation (RIP) assays were conducted with 2-week-old OsCc1::OsCRP1-GFP transgenic leaves. (a) Identification of OsCRP1 target chloroplast RNAs (cpRNAs) by RIP. cDNAs were synthesized using the immunoprecipitated RNAs and α-GFP antibodies, prior to quantitative RT-PCR. All the values were normalized based on total input RNA per sample, and bars represent the mean ± SD of four repeats. (b) Relative expression levels of cpRNAs in total RNA samples from OsCc1::OsCRP1OX, non-transgenic (NT) and OsCc1::OsCRP1RNAi plants. qRT-PCR with cDNA from NT and transgenic leaves was performed using 23 chloroplast gene-specific primer sets. All the values were normalized to the internal OsUbi1 control gene, and data bars represent the mean ± SD of two biological replicates, each of which had three technical replicates. Significant differences from the control are indicated by asterisks (Student’s t-test, * p < 0.05).