Journal: Journal of Biological Chemistry

Article Title: Long noncoding RNA Hoxb3os is dysregulated in autosomal dominant polycystic kidney disease and regulates mTOR signaling

doi: 10.1074/jbc.ra118.001723

Figure Lengend Snippet: FIGURE 2. Expression of Hoxb3os in the mouse kidney. A) Rapid amplification of cDNA ends (RACE) identifies two major Hoxb3os isoforms in the mouse kidney. Upper panel shows RNA-seq reads from a wild-type kidney visualized with IGV software (blue bars indicate reads from positive strand and red bars indicate negative strand). The two Hoxb3os isoforms (lower panel) are located 5’ to Hoxb2 and partially overlap with Hoxb3 (arrowheads indicate direction of transcription and solid rectangles indicate exons). B) Quantitative RT-PCR showing expression of Hoxb3os in different mouse organs relative to the kidney. C) Developmental expression of Hoxb3os in the embryonic (E16.5), newborn (P1) and adult (P30) mouse kidney measured by qRT-PCR. D) Percentage of Hoxb3os expression in cytoplasmic and nuclear fractions from an adult mouse kidney measured by qRT-PCR. Error bars indicate SEM.

Article Snippet: Quantitative real-time PCR- Total RNA was extracted from mouse inner medullary collecting duct (mIMCD3) cell line or mouse kidneys using RNeasy Mini Kit (Qiagen, Germantown) according to the manufacturer’s protocol. mIMCD3 cells were purchased from ATCC and were genotyped upon receipt using short tandem repeat (STR) DNA profiling. cDNA was synthesized using the iScript cDNA Synthesis Kit (Bio-Rad), and quantitative real-time PCR was performed with the iTAG Universal SYBER Green Supermix (Bio-Rad).

Techniques: Expressing, Rapid Amplification of cDNA Ends, RNA Sequencing, Software, Quantitative RT-PCR