Fig. 3
From: Single-cell CAS-seq reveals a class of short PIWI-interacting RNAs in human oocytes
The interacting protein and 3′ modification of oocyte short-PIWI-interacting RNAs (os-piRNAs). a Specificity of anti-HIWI and anti-HIWI3 antibodies. FLAG-tagged HIWI3, HIWI2, HIWI, and HILI were ectopically expressed in HEK293 cells. The cell lysates were immunoprecipitated with HIWI3- or HIWI-specific antibodies and immunoblotted with an anti-FLAG antibody. b os-piRNAs were enriched by HIWI3 immunoprecipitation (IP) in human oocytes. The abundance and length distribution of small RNAs in the input and immunoprecipitated (IP) samples of oocytes are shown. Three or four human oocytes were used for each IP reaction. Rabbit nonspecific immunoglobulin G (IgG) antibody served as a negative control. The abundance of small RNAs was normalized to an exogenous spike-in. c The ratio of small RNAs tailed with mono- and oligo-nucleotides in human and mouse oocytes is shown. The instances of 3′ adenylation in os-piRNAs are shown. d, e Composition of small RNAs in the mouse and human oocytes before and after NaIO4 oxidization. The average result of three biological replicates is shown. Source data of a–e are provided in the Source Data file
