Dataset to “novel_circ_002651 regulates the immune defense of eastern honeybee larvae against fungal invasion through sponging miR-6001-y”

This study aims to characterize the regulatory functions of a previously identified circRNA (novel_circ_002651, ac2651) and its key target miRNA (ace-miR-6001-y), in eastern honeybee worker larvae respondi ng to infection by Ascosphaera apis:
Figure 1. (A) Regulatory network between ac2651 and target miRNAs. (B) Agarose gel electrophoresis for the amplified product from ac2651. (C) Peak diagram of Sanger sequencing of the amplified fragment from ac2651, the black arrow indicates the back-splicing site. (D) Agarose gel electrophoresis for the amplified product from ace-miR-6001-y. (E) peak diagram of Sanger sequencing of the amplified fragment from ace-miR-6001-y. (F, G) Relative expression levels of ac2651 and ace-miR-6001-y in the guts of 4-, 5-, and 6-day-old larvae inoculated with A. apis.
Figure 2. (A) Loop graph of GO terms annotated by target mRNAs. (B) Loop graph of KEGG pathways annotated by target mRNAs. (C) Regulatory network between ace-miR-6001-y and immune pathway-related target DEmRNAs.
Figure 3. (A) Relative expression level of ac2651 following feeding specific siRNA. (B) Relative expression level of 3 immune genes in the larval guts after silencing ac2651. (C) Relative expression level of 3 genes relative to fungal proliferation after silencing ac2651. (D) Larval survival rate following silencing ac2651. (E) Larval chalkbrood incidence following silencing ac2651.
Figure 4. (A, F) Relative expression level of ace-miR-6001-y in the larval guts after feeding mimic and inhibitor. (B, G) Relative expression levels of 3 host immune genes following ace-miR-6001-y overexpression and knockdown. (C, H) Relative expression levels of 3 genes relevant to fungal proliferation following ace-miR-6001-y overexpression and knockdown. (D, I) Host survival rate following overexpression and knockdown of ace-miR-6001-y. (E, J) Host chalkbrood incidence following ace-miR-6001-y overexpression and knockdown.
Figure 5. (A, F) agarose gel electrophoresis for PCR amplification products. (B, G) Sanger sequencing result of the target fragment. (C, D) peak diagrams of Sanger sequencing of recombinant plasmids pmirGLO-ac2651-miR-6001-wt and pmirGLO-ac2651-miR-6001-mut. (E) Verification of binding relationship between ac2651 and ace-miR-6001-y. (H, I) peak diagrams of Sanger sequencing of recombinant plasmids pmirGLO-miR-6001-14-3-3ζ-wt and pmirGLO-miR-6001-14-3-3ζ-mut. (J) Verification of binding relationship between ace-miR-6001-y and Ac14-3-3ζ.
Figure 6. (A, B) Relative expression levels of ace-miR-6001-y and Ac14-3-3ζ after silencing ac2651. (C) Relative expression level of Ac14-3-3ζ following ace-miR-6001-y knockdown. (D) Relative expression level of Ac14-3-3ζ after simultaneously silencing ac2651 interference and knocking down ace-miR-6001-y.
Figure 7. (A) Relative expression level of 3 immune genes in the larval guts. (B) Host survival rate. (C) Relative expression levels of 3 genes related to fungal proliferation. (D) Host chalkbrood incidence.