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. 2012 Nov 29;8(11):e1003105. doi: 10.1371/journal.pgen.1003105

Figure 2. Zcchc11 deficiency does not affect quantities of mature miRNAs or miRNA–related proteins in the liver.

Figure 2

Three deep sequencing libraries were created from livers of sex- and littermate-matched 8-day-old Zcchc11+/+ or Zcchc11−/− mice. (A) Mature miRNA content, expressed as reads per million (RPM) was compared for wild type and Zcchc11-deficient livers (correlation coefficient, r = 0.975). (B) Quantitative RT-PCR was used to measure the expression of several miRNAs including some implicated in Zcchc11 pre-miRNA uridylation (Let-7), those highly expressed in the livers (miR-122), and those showing trends towards change in the deep sequencing data (miR-139 and miR-379), revealing no difference between genotypes. (C) As another approach to examining Let-7 content, Let-7a in the livers of 8-day old mice was measured by Northern blotting, and did not increase due to Zcchc11 deficiency. An adult wild type mouse was also included for comparison. (D) Immunoblots for RISC-related proteins in tissue homogenates were prepared from the livers of 8 day-old mice, showing no differences due to genotype. (E) Immunoblots for Lin-28 family members in the livers and skeletal muscles of 8 day old Zcchc11+/+ and Zcchc11−/− mice revealed no effects of Zcchc11 deficiency. Actin was measured as a loading control. For all blots, each lane represents the RNA or protein from a separate individual of the indicated age and genotype.