Altered tryptophan hydroxylase 2 expression in enteric serotonergic nerves in Hirschsprung’s-associated enterocolitis

Table 1 Details pertaining to primary and secondary antibodies used in immunofluorescence and Western blot

Host species	Monoclonal/Polyclonal	Antigen	Product code	Manufacturer	Dilution
Rabbit	Polyclonal	Tryptophan hydroxylase 2	NB-100-74555	Novus biologicals (Cambridge, United Kingdom)	1:100
Mouse	Monoclonal	TMEM-16A (ANO1)	Ab190721	Abcam (Cambridge, United Kingdom)	1:200
Mouse	Monoclonal	HuD	sc-48421	Santa-Cruz Biotechnologies (Heidelberg, Germany)	1:100
Mouse	Monoclonal	Platelet-derived growth factor alpha	sc-21789	Santa-Cruz Biotechnologies (Heidelberg, Germany)	1:100
Goat	Polyclonal	Neuronal nitric oxide synthase	ab1376	Abcam (Cambridge, United Kingdom)	1:300
Mouse	Monoclonal	Vasoactive intestinal peptide	sc-25347	Santa-Cruz Biotechnologies (Heidelberg, Germany)	1:100
Mouse	Monoclonal	Choline acetyltransferase	ab49382	Abcam (Cambridge, United Kingdom)	1:100
Mouse	Monoclonal	GAPDH	ab9484	Abcam (Cambridge, United Kingdom)	1:2000
Rabbit	Polyclonal	Mouse (secondary)	ab6728	Abcam (Cambridge, United Kingdom)	1:10000
Donkey	Polyclonal	Rabbit (secondary)	ab6802	Abcam (Cambridge, United Kingdom)	1:10000
Donkey	Polyclonal	Rabbit (Alexa Fluor® 488)	ab150073	Abcam (Cambridge, United Kingdom)	1:500
Goat	Polyclonal	Mouse (Alexa Fluor® 594)	ab150116	Abcam (Cambridge, United Kingdom)	1:500
Donkey	Polyclonal	Goat (Alexa Fluor® 555)	ab150134	Abcam (Cambridge, United Kingdom)	1:500

Table 2 Clinical details of patients with Hirschsprung’s disease included in the study

Characteristic	n = 12
Gender	Male (n = 11)	Female (n = 1)
Median age at pull-through operation	5 mo	3-14 mo
Associated syndromes	Trisomy 21	n = 5 (2 with HAEC)
Pre-operative HAEC	Yes (n = 5)	No (n = 7)
Diverting/levelling stoma	Yes (n = 3)	No (n = 9)

HAEC: Hirschsprung’s-associated enterocolitis.