Figure 1. The transgenerational memory of silencing and small RNA pools are intact in the absence of SET-2 : (A) Experimental design of RNAi induction and monitoring of transgenerational memory of silencing.(B) qRT-PCR quantification of gfp mRNA abundance in F1 wild-type and
set-2(
syb2085) animals treated for 2 generations with L4440 (empty vector) or dsRNA GFP food. Color code is the same as in (C).(C) Transgenerational scoring of silenced wild-type and
set-2(
syb2085) animals fed with L4440 or dsRNA GFP food. 3 independent wild-type and 6
set-2(
syb2085) lines were analyzed.(D) Small RNA pool sequencing in
set-2(
bn129) and
set-2(
syb2085) animals. Depleted and enriched piRNA and 22G-RNA (classified according to the nature of their genomic targets). Number of targets with significantly misregulated small RNA (
log2FC(mut vs wt) < -2 and
log2FC(mut vs wt) > 2) are indicated.(E) Classification of
set-2 enriched 22G-RNA according to their association with the 3 Argonaute proteins HRDE-1, WAGO-1 and CSR-1B in wild-type animals. Expected numbers and statistical significance were calculated using a hypergeometric assay.(F) Comparison of enriched HRDE-1 and WAGO-1 22G-RNA with genes misregulated in
set-2(
bn129) germlines. Percent of genes enriched with HRDE-1- or WAGO-1- associated 22G-RNA in
set-2 mutants and upregulated in
set-2(
bn129) germlines are indicated. Expected numbers and statistical significance were calculated using a hypergeometric assay. The list of misregulated genes in
set-2(
bn129) mutant germlines is from (Herbette et al. 2020).