Fig. 5.

uc.338 and PCBP2 are independently regulated. (A) Schematic representation of the partial exonic location of uc.338 within the PCBP2 gene. The exons of PCBP2 are indicated by dark gray boxes, and uc.338 is depicted as the light gray box. The location of the uc.338 forward (F) and reverse (R) primers used for real-time-PCR and probe used for in situ hybridization are shown. With the exception of the forward primer, these primers are located within the PCBP2 intronic region. Of the siRNAs targeting uc.338, siRNA-1 is entirely intronic, whereas siRNA-2 overlaps a few nucleotides of the coding sequence of PCBP2. F′ and R′ indicate primers used for detection of PCBP2. (B) Expression of uc.338 is plotted against PCBP2 mRNA expression in samples of normal and HCC cell lines. There is no linear correlation between uc.338 and PCBP2 gene expression (P = 0.08). (C) HepG2 cells were transfected with siRNA against PCBP2 or siRNA control. Bars represent the mean of two independent experiments performed in four replicates. *P < 0.05 compared with control siRNA. (D) The expression of uc.338 was decreased in HepG2 and Huh-7 cells by using two different siRNAs against uc.338. After 48 h, RNA was collected and uc.338 and PCBP2 expression were evaluated by real-time PCR. Bars represent the mean and SEM of two experiments performed in four replicates. *P < 0.05 relative to siRNA control. These data indicate that uc.338 does not regulate the expression of PCBP2 and that the expression of uc.338 is independent of PCBP2.