Fig. 4

SNPs nucleotide sequence variation-mediated translational inhibition of CAMTA2. a Predicted secondary structures for wild type and mutant initiation region of CAMTA2. b Schematic diagram showing the wild type and mutant containing 5′UTR reporter constructs. c HEK293 cells (3 × 10⁴ cells/well) in 96-well plates were transfected with 75 ng of purified wild-type and mutant CAMTA2 5′UTR SGFP-expressing constructs along with control RFP expressing constructs (30 ng). After 20 h, fluorescence was quantified using automated image segmentation and quantification as described in the “Methods” section. The GFP/RFP expression ratio of WT and Mut of 5′UTR are shown as mean ± SD (n = 6). ***p < 0.0001 (Student’s t test). d Real-time qRT-PCR for reporter mRNA levels. HEK293 cells were transfected with wild type and mutant CAMTA2 5′UTR reporters for 24 h. Total RNA was extracted, and qRT-PCR was performed using specific primers for SGFP and RFP as described in the “Methods” section. Data are presented as the mean ± SD of two independent experiments. **p = 0.0014 (Student’s t test)