Table 2. Biological validation of RNA-Seq results.

	Gene	qPCR (2−ddCt)			P-value	Validated?
		non-manipulated	sham-manipulated	albumen-deprived
C≠A; S≠A	SPAG4like	1.00 ± 0.27	0.56 ± 0.23	0.75 ± 0.17	NS	No
S≠A	TNFSF10	1.00 ± 0.13b	0.69 ± 0.09b	1.40 ± 0.07a	0.0006	Yes
	LAPTM4B	1.00 ± 0.16ab	1.96 ± 0.62b	0.50 ± 0.09a	0.0520	Yes
	TMEM86A	1.00 ± 0.07ab	0.93 ± 0.15b	1.42 ± 0.16a	0.0489	Yes
	CKS1B	1.00 ± 0.12ab	1.74 ± 0.39a	0.96 ± 0.08b	0.0777	Yes
	NXPH2	1.00 ± 0.08a	0.45 ± 0.07b	1.06 ± 0.20a	0.0091	Yes
	LRRC3C	1.00 ± 0.14ab	1.68 ± 0.45a	0.70 ± 0.10b	0.0776	Yes
	BMF	1.00 ± 0.10ab	0.74 ± 0.08b	1.32 ± 0.12a	0.0048	Yes
	SEMA6D	1.00 ± 0.17ab	1.70 ± 0.50a	0.63 ± 0.08b	0.0916	No
	H2B-I	1.00 ± 0.16ab	2.12 ± 0.68a	0.67 ± 0.08b	0.0681	No
	GLUL	1.00 ± 0.14	1.20 ± 0.23	1.13 ± 0.12	NS	No
	HERPUD1	1.00 ± 0.10	1.38 ± 0.36	1.17 ± 0.29	NS	No
	SLC6A6	1.00 ± 0.11	1.18 ± 0.21	1.03 ± 0.06	NS	No
	LECT2	1.00 ± 0.29	1.33 ± 0.33	0.65 ± 0.16	NS	No
	GJA1	1.00 ± 0.11	0.94 ± 0.11	0.97 ± 0.12	NS	No

Gene expression of selected DE genes (P < 0.001 and log₂-fold change >1) were measured in 8 samples per treatment via qPCR. ^a,btreatment means with different superscript are significantly different (P < 0.1).