Table 4. Hemorrhage assessment in immunocompetent C57BL/6 mice of DEN-4 2A E gene single amino acid mutant viruses.

Virus Strain	SubstitutionMutationViruses	Mouse No.	Severity of Hemorrhage		% of Hemorrhage Development
			Skin	Subcutaneous
DEN-4 2A	Wild type	1-1	+	−	20%
		1-2	−	−
		1-3	−	−
		1-4	−	−
		1-5	−	−
	E-T138P	2-1	++	−	40%
		2-2	+	−
		2-3	−	−
		2-4	−	−
		2-5	−	−
	E-G328S	3-1	−	−	20%
		3-2	−	−
		3-3	−	−
		3-4	−	−
		3-5	+	−
	E-Q438H	4-1	−	−	60%
		4-2	−	+
		4-3	−	−
		4-4	+	+
		4-5	++	−
	E-V463L	5-1	−	−	80%
		5-2	+++	−
		5-3	+	+
		5-4	+	−
		5-5	++	−

Rates represent hemorrhage percentage in each independent experiment. A three-level bleeding scoring low (“+”), medium (“++”), high (“+++”), and no bleeding (“−“) was used to indicate the severity of hemorrhaging between different experimental cases.