Action mechanism of early cerebral injuries after spontaneous subarachnoid hemorrhage by silence Ghrelin and angiogenic factor with G-patch and FHA domain 1

Figures & data

Table 1. siRNA primer information

Gene name	Direction	Sequence (5’→3’)
Ghrelin-1	Sense sequence	CCUGCUGACUUACAAAUAAAU
	Anti-sense sequence	UUAUUUGUAAGUCAGCAGGAG
Ghrelin-2	Sense sequence	GGUUCAAUGCUCCCUUCGAUG
	Anti-sense sequence	UCGAAGGGAGCAUUGAACCUG
Ghrelin-3	Sense sequence	GCUUCUGCCUCCUCUGCAACU
	Anti-sense sequence	UUGCAGAGGAGGCAGAAGCUG
Aggf1-1	Sense sequence	GGUCAAGUUCAGACUAUUACU
	Anti-sense sequence	UAAUAGUCUGAACUUGACCAA
Aggf1-2	Sense sequence	GGAAGAUGUUGGAGAAGAUGG
	Anti-sense sequence	AUCUUCUCCAACAUCUUCCGG
Aggf1-3	Sense sequence	GCAGAGUCCGUAUGUUCAAGC
	Anti-sense sequence	UUGAACAUACGGACUCUGCUG
Control	Sense sequence	UUCUCCGAACGUGUCACGUTT
	Anti-sense sequence	ACGUGACACGUUCGGAGAATT

Table 2. Evaluation standard for neurologic dysfunction

Dimensions	Specific evaluation
Diet	Ate all food (0 point); Ate some food (1 point); Not eating (2 points)
Activity	Complete free activity (0 point); Activity after being stimulated (1 point); Rare activity (2 points)
Dysfunction	No dysfunction (0 point); Unstable walking (1 point); Unable to walk (2 points)

Table 3. Information about quantitative detection primers of target genes

Gene name	Direction	Sequence (5’→3’)	Product size (bp)
Ghrelin	F	AGCCCAGCAGAGAAAGGAAT	206
	R	GAAGAAACTTTCCCAGGGCC
Aggf1	F	CAAGCTCCTTGGGCGATTTCA	161
	R	GCATAACTCCCGCTGTCTATGT
GAPDH	F	AATGGATTTGGACGCATTGGT	213
	R	TTTGCACTGGTACGTGTTGAT

Figure 1. Comparison of mRNA expression levels of target genes in rat brain tissues of each group after silence Ghrelin and Aggf1. (a showed Ghrelin mRNA expression levels, and b demonstrated Aggf1 mRNA expression levels. The comparison with sham group indicated ^aP < 0.05, the comparison with model group revealed ^bP < 0.05, and the comparison with negative group showed ^cP < 0.05).

Figure 2. Comparison of scores of rat neurologic dysfunction in each group. (The comparison with sham group indicated ^aP < 0.05, the comparison with model group revealed that ^bP < 0.05, and the comparison with negative group showed that ^cP < 0.05).

Figure 3. Comparison of rat cerebral water contents in each group. (The comparison with sham group indicated ^aP < 0.05, the comparison with model group showed ^bP < 0.05, and the comparison with negative group revealed that ^cP < 0.05).

Figure 4. Comparison of Ghrelin and Aggf1 protein expression levels of rat brains in five groups (a showed Western blotting, b displayed Ghrelin protein expression levels, and c indicated Aggf1 protein expression levels. The comparison with sham group demonstrated P < 0.05, the comparison with model group revealed ^bP < 0.05, and the comparison with negative group suggested ^cP < 0.05).

Figure 5. Comparison of p-Akt and caspase-3 expression levels of rat brains in five groups (a showed Western blotting, b displayed p-Akt protein expression levels, and c indicated caspase-3 protein expression levels. The comparison with sham group demonstrated P < 0.05, the comparison with model group revealed ^bP < 0.05, and the comparison with negative group suggested ^cP < 0.05).

Figure 6. Comparison of GSH, GSSG, and GSH/GSSG levels of rat brains in each group. (a showed the comparison of GSH contents, b demonstrated the comparison of GSSG contents, and c presented GSH/GSGG ratio values. The comparison with sham group indicated ^aP < 0.05, the comparison with model group revealed ^bP < 0.05, and the comparison with negative group demonstrated ^cP < 0.05.).