Fig. 2

Transgenic overexpression of miR-274 in the silkworm silk gland. a PCR product of the sequence containing miR-274. 1: Marker; 2: PCR product. b Verified plasmids pSL1180[Hr3/Ser1-miR-274] and pSL1180[Hr3/FibH-miR-274] by BamH I/Not I. 1: Marker; 2. Verification of pSL1180[Hr3/Ser1-miR-274]; 3–5. Verification of pSL1180[Hr3/FibH-miR-274]. c Expression cassette of miR-274 and the vector backbone piggyBac recovering.1: Marker; 2: Gel-recovered [Hr3/Ser1-miR-274]; .3: Gel-recovered [Hr3/FibH-miR-274]; 4: Gel-recovered vector backbone piggyBac[3 × P3-Red]; 5: Gel-recovered vector bankbone piggyBac[3 × P3-EGFP]. d Shecmatic diagram of MSG-specific transgenic overexpression vector of miR-274 and PSG-specific transgenic overexpression vector of miR-274. e Transgenic overexpression plasmid was verified by single enzyme digestion. 1: Marker; 2: Digested product of piggyBac[3 × P3-Red] with Asc I; 3: Digested product of [Hr3/Ser1-miR-274-SV40] with Asc I; 4: Digested product of piggyBac[3 × P3-Red, Hr3/FibH-miR-274-SV40] with Asc I. f Screening of positive individuals of miR-274-OE-MSG. (a) Observed under green light; (b) Observed under white light. g Screening of positive individuals of miR-274-OE-PSG. (a) Observed under blue light; (b) Observed under white light. h Expression of miR-274 in miR-274-OE-MSG. i Expression of miR-274 in miR-274-OE-PSG. Data represent three biological replicates with three technical replicates and are shown as mean ± SEM. *P < 0.05; ***P < 0.001