Skip to main content
. 2013 Aug 31;41(22):10086–10109. doi: 10.1093/nar/gkt777

Figure 11.

Figure 11.

Promoter RNAs link COX-2 and PLA2G4A transcription. (A) Distal and functional relationship of COX-2 and PLA2G4A. Scheme showing the location and orientation of the COX-2 and PLA2G4A promoters on chromosome 1 (top). Scheme showing the functional relationship between COX-2 and PLA2G4A within the eicosanoid signaling pathway (bottom). (B) qPCR of PLA2G4A mRNA levels after transfection of anti-miRs (42 nM) that target either miR-452 or miR-589. n = 3. (C) qPCR of PLA2G4A mRNA after transfection of mature miR-589 or miR-589 mimic (miR-589 m) (42 nM). n = 2. (D) qPCR showing effect of RNA12, RNA12nc, RNA12 or duplex RNAs (25 nM) that contain scrambled (12scr) or mismatched bases (M1, M2, M3) on expression of PLA2G4A mRNA. n = 2. (E) ChIP showing recruitment of RNAP2 at the PLA2G4A promoter following transfection with RNA12 (25 nM). n = 4. (F) qPCR showing effect of reducing promoter RNA (−81/+56) on induction of PLA2G4A mRNA by RNA12. Cells were initially transfected (TRF-1) with gapmer oligonucleotide complementary to sense RNA (gap1, 30 nM) followed by a second transfection (TRF-2) with RNA12 (25 nM). Control is a non-complementary gapmer oligonucleotide. n = 2. (G) Western showing effect of reducing cellular AGO2 protein levels on PLA2G4A protein upregulation by RNA12 (25 nM). AGO2si (12 nM) is a pool of four duplex RNAs complementary to AGO2 mRNA. Cells were first transfected (TRF-1) with mismatched control (MM) or AGO2si then transfected with MM or RNA12. (H) Western showing effect of siRNA-mediated depletion of GW182 on PLA2G4A protein levels by RNA12 (25 nM). GW182si (12 nM) is a pool of duplex RNAs complementary to mRNAs of TNRC6 paralogs. Cells were first transfected (TRF-1) with mismatched control (MM) or GW182si then transfected (TRF-2) with MM or RNA12. (I) Western showing effect of siRNA-mediated depletion of WDR5 on induction of PLA2G4A protein expression by RNA12 or RNA12nc (25 nM). WDR5si (25 nM) is a duplex RNA complementary to WDR5 mRNA. Cells were first transfected (TRF-1) with mismatched control (MM) or WDR5si then transfected (TRF-2) with MM, RNA12 or RNA12nc. n = 4. Error bars are SD. *P < 0.05, **P < 0.01 and ***P < 0.001 (t-test) relative to mismatch control.