Table 1. Gene expression changes for tsRNA target genes in S2 cells under different conditions.

Experiment	Control	No. of target genes	log2 (FCmRNA) (mean ± s.d.)	P(FCmRNA)	log2 (FCTE) (mean ± s.d.)	P(FCTE)
T3	ss-NC	396	0.067 ± 0.21	1.37 × 10−7	−0.28 ± 0.51	6.11 × 10−8
T6	ss-NC	381	0.074 ± 0.20	5.77 × 10−8	−0.14 ± 0.25	< 2.20 × 10−16
T10	ss-NC	646	0.026 ± 0.17	0.0006	−0.14 ± 0.33	< 2.20 × 10−16
AGO2 KD	ds-NC	351	0.053 ± 0.45	0.003	0.057 ± 0.35	2.22 × 10−4
Starvation	Normal	408	0.17 ± 0.55	1.91 × 10−10	−0.17 ± 0.58	7.45 × 10−11
Starvation + AGO2 KD	Starvation + ds-NC	408	0.048 ± 0.38	0.42	−0.040 ± 0.23	0.013

In total 6511 genes that were expressed in the S2 cells cultured under normal conditions were used in all the analyses.

In the T3, T6 and T10 experiments, the target mRNAs were expressed in S2 cells and had conserved 7-mer sites antisense pairing with T3, T6 and T10 tsRNAs, respectively.

In the AGO2 KD (knock-down) experiment, 351 mRNAs expressed in S2 cells had conserved target sites of AGO2-bound tsRNAs (target sites per kb >1.5).

In the serum starvation and serum starvation+AGO2 KD experiments, 408 target mRNAs expressed in S2 cells have UpSites >3.

P(FC_mRNA) and P(FC_TE) were calculated based on the comparisons (Kolmogorov–Smirnov tests) between target genes of tsRNAs and the remaining genes in the mRNA-seq and Ribo-seq data, respectively.