Fig. 4

lnc19419 regulate CPCFC which mediated malathion penetration resistance by sequestering miR-994. A Differential expression of lnc19419 in the whole body and pronotum cuticle between the MR and MS strains. Each sample of whole body and pronotum cuticle contained four flies and 20 dissected pronotum of 5-day-old adult, respectively (n = 4). B Expression of lnc19419 after injecting synthetic miR-994 or an inhibitor (n = 4). C Expression profiles of miR-994 and lnc19419 in adult flies at 3, 5, 7 and 9 days old (A3, A5, A7 and A9, n = 4). D Enrichment analysis (biotin–avidin RNA pull-down) following the binding of miR-994 to lnc19419 (n = 4). E Target site analysis, showing the interaction between miR-994 and lnc19419. Mutated-lnc19419 contained the sequence 5′-AAAAGAA-3′ (red). F Luciferase activity indicating the targeting of miR-994 by lnc19419 in vitro. lnc19419-wt and lnc19419-mut indicate HEK293-T cells containing the 3′ UTR of wild-type and mutated lnc19419, respectively (n = 4). G CPCFC and lnc19419 expression levels after silencing lnc19419.(n = 4). H The relative CPCFC content of pronotum cuticular sections after silencing lnc19419. CPCFC protein expression was determined by the fluorescence signal intensity over length in each sample (n = 6). I Analysis of pronotum cuticular ultrastructure after silencing lnc19419. J Analysis of the cuticular thickness after silencing lnc19419 (n = 10). Each thickness value is the mean of five randomly selected cross-sectional measurements. K The malathion penetration rate after silencing lnc19419. Each eluant contained 30 5-day-old adult flies (n = 3). L The effect of lnc19419 silencing on susceptibility to malathion. Survival rate was observed at 0–3 day after malathion exposure (n = 126)