Figure 1 | Oncogene

Figure 1

From: Genome-wide loss-of-function genetic screening identifies opioid receptor μ1 as a key regulator of L-asparaginase resistance in pediatric acute lymphoblastic leukemia

Figure 1

Genome-wide RNAi screening identifies loss of OPRM1 in the development of L-asparaginase resistance in leukemic cells. (a) Reduced L-asparaginase sensitivity in shRNA library-infected cells (•) compared to parental cells (○) and cells infected with an empty pRS vector (▪). Cells were treated with increasing concentrations of L-asparaginase for 4 days and viability was measured. Values are means±s.d. of three independent experiments. (b) Presence of shRNA inserts in cells infected with retrovirus carrying shRNA library. PCR was performed using gDNAs as template and pRS forward (5'-CCCTTGAACCTCCTCGTTCGACC-3') and reverse (5'-GAGACGTGCTACTTCCATTTGTC-3') primers. (c) Identification of genes responsible for L-asparaginase resistance. Ten L-asparaginase-resistant clones isolated by soft agar colony formation assay were subjected to gDNA isolation and PCR using the primers described in b. 642 bp PCR products resolved in 1% agarose gel were cut, extracted and sequenced using pRS-sequence primer (5'-GCTGACGTCATCAACCCGCT-3').

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