Skip to main content
. Author manuscript; available in PMC: 2015 Oct 1.
Published in final edited form as: Nat Struct Mol Biol. 2015 Mar 2;22(4):319–327. doi: 10.1038/nsmb.2982

Figure 5. Ectopically expressed lnc-pri-miR-122 switches to CPA when Microprocessor activity is inhibited.

Figure 5

a. Schematic of lnc-pri-miR-122 expression construct driven under HIV-LTR promoter, with locations of northern probe, PCR amplicon and PAS (pA1). WT and Δ plasmids were generated with and without the pre-miR-122 hairpin. b. Northern blot showing that mature miR-122 is expressed from the WT, but not Δ, plasmid following transfection into HeLa cells. U6 snRNA is loading control. c. Northern blot showing spliced and unspliced lnc-pri-miR-122 transcripts generated from the WT plasmid transfected into HeLa cells are the same size as endogenous transcripts in Huh7 cells. d. Northern blot showing spliced and unspliced lnc-pri-miR-122 transcripts generated from the Δ plasmid are larger than the WT transcripts upon transfection into HeLa cells. e. Northern blot showing transcripts generated from the WT plasmid increase in size following Drosha or DGCR8 depletion in HeLa cells. f. RT-qPCR analysis of pA+ or pA− fractionated RNA extracted from siRNA-treated HeLa cells transfected with the WT plasmid. pA+ RNA levels of U6 snRNA, GAPDH mRNA and lnc-pri-miR-122 are expressed relative to pA− which were set to 1. Error bars represent s.d. of an average (n=3 independent experiments). g. Northern blot showing that mutation of PAS (pA1) does not affect migration of WT lnc-pri-miR-122, but leads to loss of unspliced and spliced transcripts generated from the Δ plasmid upon transfection into HeLa cells.