These procedures were performed as described previously63 (link), 64 (link). Primers used in RT-qPCR are indicated in Supplementary Table 1 . The miR-122, miR-16 (as internal control for vesicular miR-122), and U6 primers (as internal control for intracellular miR-122) were purchased from Qiagen. Determination of PKM isoforms was performed as described65 (link). Products of RT-PCR were digested with NcoI, PstI, or both enzymes, plus an uncut control, and separated on an agarose gel with Sybr safe. The presence of a PstI digestion site indicates the splicing isoform M2 whereas the NcoI site indicates isoform M1.
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Quantifying Cellular miRNA and Isoform Expression
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Corresponding Organization :
Other organizations : City of Hope, University of California, Riverside, City Of Hope National Medical Center, Tianjin Medical University Cancer Institute and Hospital
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Variable analysis
independent variables
- Primers used in RT-qPCR
- Expression of miR-122, miR-16, and U6
- PKM isoforms
- Procedures performed as described previously
- MiR-16 (as internal control for vesicular miR-122)
- U6 (as internal control for intracellular miR-122)
- Uncut control for RT-PCR products
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