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Briefly, following the applied treatment, cells were collected by trypsinization, washed, and incubated with 0.3% formaldehyde and glycine (0.125 M), and cell pellets were re-suspended in the RIP buffer as previously described22. Lysates were then incubated with the anti-IGF2BP1 antibody (Santa Cruz Biotech). IGF2BP1-bound pellets were washed, re-suspended and incubated with three times in cold PBS, and re-suspended and incubated with the proteinase K-containing buffer containing. IGF2BP1-bound RNA was isolated. LIN28B-AS1 expression was tested by qPCR.
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