Figure 2
The 5′UTR of NRXN2α represses protein expression of downstream cistron. (A) Schematic diagram of plasmid constructs. The horizontal line represents the 5′ UTR of NRXN2α; the black rectangles indicate the open reading frame of firefly luciferase (Fluc) and the open rectangles indicate the open reading frame of human NRXN2α gene. (B) SH-SY5Y cells were co-transfected with luciferase reporters and p3PRluc. Firefly luciferase activity was measured 24 h after transfection, and Renilla luciferase activity was used to normalize for transfection efficiency. Data are shown as relative values over the samples transfected with empty control p4P plasmid. (C) Western Blot of luciferase protein. HEK293 cells were transiently co-transfected with luciferase reporters and EGFP expression plasmid. (D) Density analysis of the luciferase protein. The density was normalized to that of β-actin. (E) Quantification of the firefly luciferase mRNAs in SH-SY5Y cells transfected with reporter plasmids. The luciferase mRNA levels were normalized to β-actin mRNA levels. (F) Quantification of the NRXN2α mRNAs in HEK293 cells transfected with NRXN2α expression plasmids. The NRXN2α mRNA levels were normalized to β-actin mRNA levels. (G) Western Blot of neurexin2α protein expression. HEK293 cells were transiently co-transfected with NRXN2α and EGFP expression plasmid. (H) Density analysis of the neurexin2α protein. The density was normalized to that of β-actin. All values were expressed as means ± S.D., N = 3. ***P < 0.001 and **P < 0.01 by the Student’s t test. The full-length blots for (C,G) are presented in Supplementary Fig. 1.
