Immune and microRNA responses to Helicobacter muridarum infection and indole-3-carbinol during colitis

Table 1 Primers used for transcription analysis

Gene	Forward primer (5’-3’)	Reverse primer (5’-3’)
Actb (β-actin)	TCACCCACACTGTGCCCATCTACG	CAGCGGAACCGCTCATTGCCAATGG
Il17a	TCAGCGTGTCCAAACACTGAG	CGCCAAGGGAGTAAAGACTT
Foxp3	AGCAGTCCACTTCACCAAGG	GGATAACGCCAGAGGAGCTG
Rorc	CCGCTGAGAGGGCTTCAC	TGCAGGAGTAGGCCACATTACA
Il10	TGAATTCCCTGGGTGAGAAGC	ATCACTCTTCACCTCCAC
Il6	AGCCAGAGTCCTTCAGAGAGAT	AAAAAGTGCCGCTACCCTGA

Table 2 Colitis symptom score P values

Treatment groups2		% Weight	Stool blood	Diarrhea	DAI
Ctrl vs	Hm	0.2495	0.61421	0.02481	0.06591
	Hm/DSS	< 0.00011	< 0.00011	< 0.00011	< 0.00011
	Hm/DSS/I3C	0.9574	< 0.00011	< 0.00011	< 0.00011
	DSS	0.0552	< 0.00011	< 0.00011	< 0.00011
	DSS/I3C	0.1381	< 0.00011	< 0.00011	< 0.00011
Hm vs	DSS	0.9823	< 0.00011	< 0.00011	< 0.00011
	Hm/DSS	< 0.00011	< 0.00011	< 0.00011	< 0.00011
Hm/DSS vs	DSS	< 0.00011	0.00071	0.00411	< 0.00011
	Hm/DSS/I3C	< 0.00011	< 0.00011	< 0.00011	< 0.00011
DSS vs	DSS/I3C	0.9977	0.00261	< 0.00011	< 0.00011

¹Significant P values.

²Ctrl: Control; Hm: Helicobacter muridarum; Hm/DSS: Helicobacter muridarum plus DSS; Hm/DSS/I3C: Helicobacter muridarum plus DSS plus I3C; DSS/I3C: DSS plus I3C.

Table 3 MicroRNA expression

	Raw expression data						Fold changes
miRNA	Ctrl	Hm	Hm/DSS	DSS/Hm/ I3C	DSS	DSS/ I3C	Hm vs Ctrl	Hm/DSS vs Ctrl	DSS/Hm/I3C vs Ctrl	DSS vs Ctrl	DI vs Ctrl	DMI vsDHM	DI vs DSS
mmu-let-7a-5p	12.93	11.14	5.72	9.11	10.71	10.59	-3.45	-147.58	-14.06	-4.65	-5.07	10.5	-1.09
mmu-let-7b-5p	14.14	11.84	7.97	10.45	11.21	10.89	-4.92	-72.01	-12.96	-7.62	-9.56	5.56	-1.25
mmu-let-7d-5p	13.76	11.52	8.09	10.44	11.1	10.86	-4.71	-50.67	-9.97	-6.29	-7.42	5.08	-1.18
mmu-let-7e-5p	11.06	10.06	1.7	8.39	9.6	9.23	-2	-654.64	-6.34	-2.74	-3.54	103.31	-1.29
mmu-let-7g-5p	10.31	6.71	1.37	2.01	7.52	6.16	-12.19	-491.93	-317.1	-6.92	-17.81	1.55	-2.58
mmu-miR-103-3p	11.85	10.44	6.42	10.09	10.88	10.02	-2.66	-43.07	-3.39	-1.96	-3.57	12.7	-1.82
mmu-miR-106a-5p	9.29	5.45	1.15	2.03	5.51	2.78	-14.34	-282.96	-153.91	-13.78	-91.54	1.84	-6.64
mmu-miR-127-3p	7.9	4.55	0.95	2.8	2.43	1.91	-10.18	-123.98	-34.2	-44.34	-63.55	3.63	-1.43
mmu-miR-128-3p	5.81	0.79	0.94	1.04	0.77	1.3	-32.4	-29.23	-27.31	-33.04	-22.87	1.07	1.44
mmu-miR-135a-1-3p	3.64	1.3	1.8	1.26	1.43	2.12	-5.04	-3.57	-5.19	-4.62	-2.86	-1.45	1.62
mmu-miR-140-3p	10.2	9.92	1.83	7.91	9.62	8.3	-1.21	-329.16	-4.87	-1.49	-3.73	67.62	-2.51
mmu-miR-140-5p	6.33	1.38	0.79	1.02	1.38	0.87	-30.86	-46.56	-39.48	-30.86	-43.84	1.18	-1.42
mmu-miR-142-5p	2.9	0.99	1.35	0.99	1.16	1.01	-3.78	-2.93	-3.78	-3.34	-3.72	-1.29	-1.11
mmu-miR-145a-5p	12.99	11.51	6.79	10.2	11.15	10.28	-2.78	-73.5	-6.89	-3.57	-6.55	10.66	-1.83
mmu-miR-146a-5p	10.34	7.84	1.24	3.54	7.72	7.84	-5.65	-550.48	-111.6	-6.13	-22.7	4.93	-3.7
mmu-miR-150-5p	13.4	11.1	4.14	8.96	10.4	9.76	-4.92	-614.48	-21.78	-8.01	-12.51	28.22	-1.56
mmu-miR-155-5p	9.86	8.15	1.54	3.58	7.65	5.31	-3.27	-320.03	-77.72	-4.61	-23.48	4.12	-5.09
mmu-miR-15b-5p	11.17	10.17	1.48	8	9.92	8.84	-1.99	-824.5	-9.01	-2.37	-5.02	91.56	-2.12
mmu-miR-16-5p	13.18	10.38	1.35	9.1	10.8	9.11	-7	-3646.25	-16.92	-5.22	-16.9	215.51	-3.23
mmu-miR-17-5p	10.71	9.01	1.33	7.8	9.29	7.82	-3.25	-666.3	-7.53	-2.67	-7.43	88.54	-2.78
mmu-miR-185-5p	9.27	6.58	0.99	5.64	5.86	5.07	-6.45	-310.71	-12.33	-10.58	-18.36	25.19	-1.74
mmu-miR-18a-5p	7.32	1.37	0.84	1.37	1.41	1.74	-61.84	-89.41	-61.84	-60.29	-47.91	1.45	1.26
mmu-miR-195a-5p	10.59	8.03	1.91	5.03	8.22	7.39	-5.89	-409.38	-47.19	-5.17	-9.14	8.68	-1.77
mmu-miR-196b-5p	3.11	0.94	0.94	1.08	1.05	0.83	-4.49	-4.49	-4.09	-4.19	-4.86	1.1	-1.16
mmu-miR-199a-5p	9.5	4.56	1.77	2.01	7.72	1.77	-2.03	-101.88	-21.49	-3.71	-78.21	4.74	-21.08
mmu-miR-19a-3p	4.08	1.56	1.39	0.91	1.24	1.25	-5.73	-6.45	-9.02	-7.15	-7.13	-1.4	1
mmu-miR-19b-3p	9.43	6.13	1.18	5.87	7.97	5.61	-9.84	-303.99	-11.79	-2.74	-14.07	25.78	-5.14
mmu-miR-20b-5p	9.03	4.87	1.76	2.29	5.85	1.96	-17.77	-154.33	-106.75	-9.05	-133.96	1.45	-14.8
mmu-miR-221-3p	9.19	6.07	1.24	1.42	7.45	5.25	-8.65	-246.56	-218.06	-3.34	-15.34	1.13	-4.59
mmu-miR-222-3p	8.89	7.65	1.12	4.11	6.24	4.58	-2.36	-219.12	-27.5	-6.29	-19.86	7.97	-3.16
mmu-miR-223-3p	5.45	1.57	1.01	0.99	1.84	0.87	-14.69	-21.58	-21.99	-12.14	-23.85	-1.02	-1.96
mmu-miR-24-2-5p	12.74	10.84	1.78	9.51	10.8	10.12	-56.17	-101.73	-64.83	-72.38	-48.35	1.57	1.5
mmu-miR-27a-3p	9.96	5.94	1.35	1.57	8.22	5.2	-16.28	-390.62	-337.26	-3.34	-27.11	1.16	-8.12
mmu-miR-28a-3p	7.53	4.75	0.83	1.4	1.49	1.12	-6.86	-103.85	-69.7	-65.83	-84.8	1.49	-1.29
mmu-miR-29a-3p	10.97	8.84	2.38	7.73	9.45	7.38	-4.37	-386.19	-9.45	-2.86	-12.06	40.85	-4.21
mmu-miR-29c-3p	5.38	0.87	1.23	1.28	2.05	1.8	-22.8	-17.81	-17.22	-10.08	-12.01	1.03	-1.19
mmu-miR-30e-5p	7.8	1.13	1.64	1.64	3.46	1.32	-101.76	-71.46	-71.46	-14.68	-89.04	1	-4.42
mmu-miR-345-5p	5.41	2.46	2.85	3.65	2.81	1.18	-7.76	-5.93	-3.41	-6.1	-18.84	1.74	-3.09
mmu-miR-374b-5p	4.34	0.92	1.19	0.89	1.58	1.13	-10.7	-8.87	-10.91	-6.78	-9.23	-1.23	-1.36
mmu-miR-423-5p	7.07	3.13	1.29	4.25	2.56	1.26	-15.33	-54.69	-7.03	-22.66	-55.78	7.78	-2.46
mmu-miR-491-5p	2.95	1.39	1.15	1.01	1.34	1.07	-2.96	-3.49	-3.84	-3.06	-3.7	-1.1	-1.21
mmu-miR-532-5p	8.4	5.55	0.89	3.43	3.45	1.75	-7.22	-183.34	-31.45	-30.94	-101.04	5.83	-3.27
mmu-miR-760-3p	2.44	0.99	0.74	1.1	1.15	1.17	-2.73	-3.25	-2.54	-2.46	-2.42	1.28	1.02
mmu-miR-877-5p	4.05	3.56	1.93	2.14	3.28	1.8	-1.4	-4.35	-3.75	-1.7	-4.76	1.16	-2.79
mmu-miR-93-5p	10.41	8.67	1.86	7.64	8.66	6.85	-3.34	-373.43	-6.79	-3.35	-11.77	54.98	-3.52

Ctrl: Control; Hm: Helicobacter muridarum; Hm/DSS: Helicobacter muridarum plus DSS; Hm/DSS/I3C: Helicobacter muridarum plus DSS plus I3C; DSS/I3C: DSS plus I3C.