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. 2019 Jun 5;17(6):e3000309. doi: 10.1371/journal.pbio.3000309

Fig 8. Somatic AMPK activity is sufficient to restore germ cell quiescence and integrity through the transmission of small RNAs in aak(0) mutants.

Fig 8

(A) Plasmid constructs that contain aak-2 cDNA driven by tissue-specific promoters were injected into daf-2;aak(0) mutants, and both the dauer-dependent germline hyperplasia and the PD sterility were evaluated for each transgenic strain. All transgenic lines are extrachromosomal and are represented by square brackets, and 3 independently generated lines were used for quantification. PD fertility was assessed 24 h following the temperature shift after animals were maintained minimally 24 h in dauer. *P < 0.05 when compared to [unc-119p::aak-2]; ***P < 0.0001 and **P < 0.001 using Marascuilo procedure when compared to daf-2; aak(0); n = 50. (B) Whole-animal DAPI staining was performed to quantify the number of germ cells present in the dauer gonad in the transgenic lines and compared to controls. ***P < 0.0001 and **P < 0.001 using the two-tailed t test when compared to daf-2; aak(0); n = 50. (C, D) All the analyzed images are merged, condensed Z stacks. The graphs represent the average immunofluorescence for H3K4me3 and H3K9me3 normalized to DAPI across the dissected gonad. **P < 0.001 using F-test of variance when compared to daf-2; aak(0), and n = 10. (E) Disrupting soma-to-germline transmission of dsRNA by compromising the function of sid-1 partially restores fertility in the daf-2; aak(0) PD animals. A number of animals laying eggs were counted, and the mean is shown ± SD. *P < 0.05 using Marascuilo procedure when compared to daf-2; aak(0), and n = 100. Underlying data can be found in S1 Data. aak, AMP-activated Protein Kinase subunit; AMPK, AMP Kinase; ckb, Choline Kinase B; DAF, DAuer Formation abnormal; dpy, DumPY; dsRNA, double-stranded RNA; Erythroid-Like Transcription factor family; H3K4me3, histone H3 lysine 4 trimethylation; H3K9me3, histone H3 lysine 9 trimethylation; No., number; PD, post-dauer; RNAi, RNA interference; sid-1, Systemic RNAi Defective; sulp, SULfate Permease family; sur-5, SUppressor of activated let-60 Ras; unc, UNCoordinated.