Fig. 3: Effect of silencing MMEJ repair pathway-related genes on eccDNA^fib-L formation.

A, C, E, G The interference efficiency of siRNAs in BmN cells. 1 × 10⁵ BmN cells were cultured for 24 h. Then, cells were transfected with siRNA at the final concentration of 5 µmol/mL. The total RNA was extracted at 48 h post transfection and reversely transcribed into cDNA. The relative expression levels of genes related to MMEJ were determined by qPCR. Considering the NC-siRNA as the control and the TIF-4A gene as the internal reference. B, D, F, H The effect of silencing MMEJ repair pathway-related genes on eccDNA^fib-L formation in BmN cells. The transfected cells with siRNAs were mentioned above. The cells were collected at 48 h post transfection for DNA extraction. The efficiency of eccDNA^fib-L formation was determined by qPCR. I, K, M, O Interference efficiency of siRNA in silkworm. siRNA (5 µmol/mL, 5 µL) was injected into silkworm larvae of the 5th instar. The total RNA of the silk gland was collected at 72 h post-injection, and reversely transcribed into cDNA. The relative expression levels of genes related to MMEJ were determined by qPCR. The NC-siRNA was used as a control and TIF-4A gene was used as an internal reference. J, L, N, P The effect of silencing MMEJ repair pathway-related genes on eccDNA^fib-L formation in silkworm. The silk glands of injected silkworms injected with siRNAs were collected at 72 h post-injection to extract DNAs. Using eccDNA^fib-L-junction primer, the efficiency of eccDNA^fib-L formation was determined by qPCR (n = 3).