Figure S7: Expression of miR-21 isoforms in the knockdown and overexpression experiments.
(A-J) The expression levels of miR-21 isomiRs were normalized to the expression level of miR-21+C. Each panel shows the change in the normalized isomiR expression relative to the average of the normalized expression in the corresponding NC condition. In A-C and E-J, miR-21 isomiRs are denoted by their 3’ terminal nucleotide. I and J refer to the same experiment, in which overexpression of PAPD5 in MCF7 cells by Lentivirus transduction was followed by knockdown of PARN by siRNA. (A) Expression of miR-21 isomiRs as measured by short RNA sequencing upon knockdown of PAPD5 in THP1 cells (two replicates). (B) Expression of miR-21 isomiRs as measured by short-RNA sequencing upon overexpression of PAPD5 in THP1 cells. (C) Expression of miR-21 isomiRs as measured by short-RNA sequencing upon overexpression (OE) of PAPD5 in MCF7 cells, sampled two weeks after PAPD5 transduction (three replicates). (D) Expression of the 22-nt miR-21 isomiR, normalized to the expression of the miR-21+C isomiR, upon overexpression of PAPD5 in MCF7 cells (three replicates), measured by isomiR-specific qPCR. (E) Expression of miR-21 isomiRs as measured by short-RNA sequencing upon overexpression of PAPD5 in MCF7 cells, sampled 8 days after PAPD5 transduction (three replicates). (F) Expression of miR-21 isomiRs as measured by short-RNA sequencing upon knockdown of PARN in THP1 cells (two replicates). The second replicate showed an outlier in the expression of the 20 nt degradation product. This outlier was not observed in the first replicate, nor in the three replicates for PARN knockdown in MCF7 cells (H). (G) Expression of miR-21 isomiRs as measured by short-RNA sequencing upon knockdown of PAN3 in THP1 cells (two replicates). (H) Expression of miR-21 isomiRs as measured by short RNA sequencing upon knockdown of PARN in MCF7 cells (three replicates). (I) Expression of miR-21 isomiRs as measured by short-RNA sequencing upon overexpression of PAPD5 followed by knockdown of PARN in MCF7 cells (three replicates), normalized to PAPD5 OE control to show the effect of PAPD5 overexpression. (J) Expression of miR-21 isomiRs as measured by short-RNA sequencing upon overexpression of PAPD5 in MCF7 cells followed by knockdown of PARN (three replicates), normalized to PARN KD control to show the effect of PARN knockdown.