Characterization and comparison of human sperm sncRNA signatures between traditional sncRNA-seq and PANDORA-seq (IMAGE)

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Characterization and comparison of human sperm sncRNA signatures between traditional sncRNA-seq and PANDORA-seq

Caption

(A) Comparison of the relative ratio of clean reads to raw reads and reads aligned to the genome to clean reads between traditional sncRNA-seq and PANDORA-seq. (B) The scatter plots depicting the correlation of miRNA, tsRNA, and rsRNA expression profiles of paired traditional sncRNA-seq and PANDORA-seq. Spearman correlation coefficients and P-values are shown. (C) Comparison of the relative expression levels of four major sncRNA origins (miRNA, tsRNA, rsRNA, ysRNA, and piRNA) in healthy human sperm determined by the PANDORA-seq protocol. (D) Expression levels of miRNAs, tsRNAs, and rsRNAs in human sperm samples validated by northern blotting. (E) The radar plot showing the different relative expression proportions of each tsRNA subcategory with respect to the two protocols. (F) Northern blotting validation of tsRNALeu(TAG), tsRNALys(TTT), and tsRNAGly(GCC) in human sperm. (G) Visualization of the proportional distribution of cyto-tsRNAs and mt-tsRNAs in healthy human sperm determined by the PANDORA-seq protocol. (H) The radar plots illustrating the relative expression proportions of each tsRNA subcategory in relation to three distinct tsRNA origins (5′, inner′, and 3′). (I) Sequence mapping location (left), expression profile (middle), and reverse transcription PCR validation (right) of tsRNAAla(AGC), tsRNAArg(TCG), and tsRNAGlu(CTC) in human sperm. (J) Distribution of expression proportions for six nucleus-encoded rsRNAs and two mitochondria-encoded rsRNAs. (K) The boxplots showing the relative expression of five nucleus-encoded rsRNAs and two mitochondria-encoded rsRNA categories. (L) Visualization of expression signature and sequence mapping location of rsRNA-18S and rsRNA-28S in human sperm samples. (M) Northern blotting validation of rsRNA-5.8S, selected rsRNA-18S main peaks (peak #1, peak #2), and selected rsRNA-28S main peaks (peak #1, peak #2) in human sperm samples.

Credit

Ruofan Huang, Yiting Yang, Wenlin Jiang, Zheng Cao, Junchao Shi, Xiao-Ou Zhang, Yunfang Zhang

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