Supporting Information

Table S1. Number of sequences analyzed, operational taxonomic units (OTUs), estimated

OTU richness (abundance-based coverage estimator [ACE] and Chao1), and coverage.

Sample Valid reads OTUs Ace Chao1 Goods Lib. CoverageNOR_1 2413 322 445 471 0.95NOR_2 2472 401 559 559 0.94NOR_3 2324 368 652 569 0.93NOR_4 2652 366 529 573 0.95NOR_5 2151 271 414 414 0.95

Mean±S.D. 2402±165 a 346±45 520±85 517±64 0.94±0.01　 　 　 　 　 　

Sample Valid reads OTUs Ace Chao1 Goods Lib. CoverageTNBS_1 1754 293 601 499 0.93TNBS_2 2111 289 586 450 0.94TNBS_3 2075 348 730 606 0.92TNBS_4 2230 321 686 548 0.93TNBS_5 3088 511 976 826 0.93

Mean±S.D. 2252±447 352±82 716±141 586±146 0.93±0.01　 　 　 　 　 　

Sample Valid reads OTUs Ace Chao1 Goods Lib. CoverageEC_1 1931 397 1016 703 0.90EC_2 2109 395 751 656 0.92EC_3 1941 326 677 551 0.92EC_4 1933 417 924 690 0.90EC_5 2096 438 1186 856 0.89

Mean±S.D. 2002±75 394±42 911±204 691±110 0.905222　 　 　 　 　 　

Sample Valid reads OTUs Ace Chao1 Goods Lib. CoverageEC+LJ_1 2256 293 420 488 0.95EC+LJ_2 1453 291 572 471 0.91EC+LJ_3 3079 376 622 551 0.96EC+LJ_4 3232 341 484 479 0.96EC+LJ_5 2690 411 576 580 0.94Mean±S.D. 2542±641 342±47 535±73 514±43 0.94±0.002

aValues indicate mean± S.D. (n=5). #P<0.05 vs normal control group (NOR). *P<0.05 vs EC-

treated control group (EC).

Table S2. Difference in the composition (percent of total sequences) of fecal bacterial genus 1

isolated from normal control, TNBS-, EC-, and EC plus LJ-treated samples.

GenusComposition (%)

NOR TNBS EC EC+LJKE159538_g 18.69 ± 10.98a 0.27 ± 0.23# 1.91 ± 0.95# 4.48 ± 3.98DQ815871_g 9.38 ± 11.77 4.94 ± 2.23 22.65 ± 3.75# 7.47 ± 1.99*

Alistipes 12.76 ± 6.87 3.81 ± 2.82 3.73 ± 0.73# 12.97 ± 2.17*

EF602759_g 1.15 ± 0.89 8.11 ± 3.64# 4.73 ± 1.85# 2.15 ± 3.03EF603735_g 0.00 ± 0.00 8.67 ± 4.33# 3.63 ± 2.38# 1.75 ± 1.41Bacteroides 1.22 ± 1.23 5.96 ± 1.29# 2.53 ± 0.84# 4.32 ± 2.24Oscillibacter 3.32 ± 2.48 1.95 ± 1.37 1.16 ± 0.45 7.32 ± 3.31*

Prevotella 3.86 ± 2.70 1.07 ± 0.86 3.89 ± 2.00 0.87 ± 1.26Pseudoflavonifractor 3.32 ± 1.17 2.19 ± 0.77 1.71 ± 0.64 6.02 ± 2.05*

AB626958_g 6.61 ± 4.27 1.66 ± 1.26# 1.79 ± 0.80# 2.65 ± 1.81Hungatella 4.55 ± 2.66 0.46 ± 0.31# 0.85 ± 0.48# 2.27 ± 0.75*

EU939387_g 0.18 ± 0.29 9.90 ± 9.32 0.00 ± 0.00 0.00 ± 0.00AB606322_g 1.37 ± 1.34 1.38 ± 0.85 2.64 ± 0.83 3.47 ± 1.85Helicobacter 0.68 ± 0.57 3.80 ± 1.51# 2.92 ± 2.51 0.61 ± 0.39EF406806_g 1.57 ± 1.39 2.99 ± 1.19 1.92 ± 0.53 0.24 ± 0.19*

HM124280_g 0.98 ± 0.80 2.96 ± 0.72# 2.12 ± 0.81 0.43 ± 0.63*

AB626939_g 3.08 ± 4.64 0.23 ± 0.30 0.26 ± 0.24 2.75 ± 1.63*

FJ880046_g 0.80 ± 0.78 1.08 ± 0.60 3.51 ± 1.26# 0.81 ± 0.49*

FJ880859_g 0.32 ± 0.20 1.35 ± 1.17 3.79 ± 1.24# 0.52 ± 0.42*

AY239469_g 0.51 ± 0.34 1.02 ± 0.48# 3.31 ± 1.25# 0.76 ± 0.77*

EF602759_f_uc 0.36 ± 0.31 1.39 ± 0.44# 3.52 ± 1.28# 0.27 ± 0.10*

EU509860_g 0.30 ± 0.39 0.60 ± 0.44 0.45 ± 0.25 3.54 ± 3.16Parabacteroides 0.63 ± 1.00 2.31 ± 0.85# 0.75 ± 0.36 0.65 ± 0.76EU622683_g 1.11 ± 0.74 0.49 ± 0.50 1.49 ± 1.09 2.15 ± 0.93aValues indicate mean± S.D. (n=5). #P<0.05 vs normal control group (NOR). *P<0.05 vs EC-

treated control group (EC).

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Table S3. Difference in the composition (percent of total sequences) of fecal bacterial species

isolated from normal control, TNBS-, EC-, and EC plus LJ-treated samples.

SpeciesComposition (%)

Nor TNBS EC EC+LJAB606279_s 6.10 ± 8.34a 0.12 ± 0.12 0.46 ± 0.44 2.31 ± 1.804P003085_s 0.00 ± 0.00 7.61 ± 4.08# 6.05 ± 3.97# 0.00 ± 0.00*

DQ815759_s 5.78 ± 3.50 0.54 ± 1.04# 0.19 ± 0.15# 4.37 ± 3.85*

AM238019_s 0.01 ± 0.02 9.71 ± 9.21 0.00 ± 0.00 0.00 ± 0.00Bacteroides acidifaciens 1.06 ± 1.05 4.75 ± 1.20# 2.50 ± 2.60 0.83 ± 0.42EF603701_s 2.90 ± 3.52 1.56 ± 1.33 1.86 ± 0.57 3.09 ± 1.22*

AB606322_s 1.36 ± 1.34 1.35 ± 0.83 2.53 ± 0.89 3.44 ± 1.86DQ815942_s 3.71 ± 2.60 0.55 ± 0.96# 0.00 ± 0.00# 0.87 ± 1.26EF605065_s 7.57 ± 7.63 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00EF603109_s 1.54 ± 1.85 1.07 ± 0.39 3.70 ± 1.46# 0.15 ± 0.17*

JQ085133_s 0.01 ± 0.02 4.54 ± 3.96# 1.66 ± 0.91# 0.00 ± 0.00*

DQ815748_s 0.48 ± 0.55 0.05 ± 0.10 0.02 ± 0.03 4.91 ± 3.49*

EF406536_s 0.78 ± 0.76 1.06 ± 0.59 2.93 ± 0.68# 0.81 ± 0.49*

EF602759_f_uc_s 0.36 ± 0.31 1.39 ± 0.44# 3.83 ± 0.88# 0.27 ± 0.10*

EF406459_s 0.03 ± 0.06 0.27 ± 0.28 5.13 ± 3.07# 0.43 ± 0.38*

EF406443_s 3.34 ± 5.07 0.20 ± 0.23 1.47 ± 0.82 0.57 ± 0.77Helicobacter ganmani 0.00 ± 0.00 3.09 ± 1.51# 2.33 ± 2.74 0.00 ± 0.00DQ815871_g_uc 0.64 ± 0.82 0.40 ± 0.19 3.77 ± 0.69# 0.47 ± 0.25*

DQ815395_s 0.37 ± 0.33 0.35 ± 0.28 1.39 ± 0.65# 2.68 ± 0.55EF406712_s 1.65 ± 1.85 1.17 ± 0.45 1.68 ± 0.53 0.40 ± 0.28EF406456_s 0.97 ± 0.70 0.45 ± 0.47 1.23 ± 0.75 1.95 ± 0.88FJ881271_s 2.72 ± 4.40 0.10 ± 0.10 0.03 ± 0.04 1.28 ± 0.93*

AB606321_s 2.74 ± 3.35 0.58 ± 0.29 0.59 ± 0.45 0.78 ± 0.36EF406806_s 0.67 ± 0.79 2.18 ± 0.86# 0.92 ± 0.46 0.04 ± 0.07*

aValues indicate mean± S.D. (n=5). #P<0.05 vs normal control group (NOR). *P<0.05 vs EC-

treated control group (EC).

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Figure S1. 2,4,6-Trinitrobenzensulfonic acid (TNBS) caused colitis and memory impairment

in mice. Colitis markers NF-κB activation and iNOS, COX-2, and tight-junction protein

expression (a) were measured in the colon. BDNF expression and NF-κB activation were

measured in the hippocampus (b). Proteins were measured by immunoblotting. Values

indicate mean± S.D. (n=10). *P<0.05.

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Figure S2. Escherichia coli (EC) caused colitis and memory impairment in mice. Colitis

markers including NF-κB activation, iNOS and COX-2 expression (a), and tight-junction

protein expression (b) were measured in the colon. BDNF expression and NF-κB activation

was measured in the hippocampus (l). Proteins were measured by immunoblotting. Values

indicate mean± S.D. (n=10). *P<0.05.

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Figure S3. Lactobacillus johnsonii (LJ) attenuated 2,4,6-trinitrobenzensulfonic acid (TNBS)-

induced colitis and memory impairment. BDNF expression and NF-κB activation were

measured in the hippocampus (a). Colitis markers including NF-κB activation and iNOS and

COX-2 expression were measured in the colon (b). Proteins were measured by

immunoblotting. Values indicate mean± S.D. (n=10). *P<0.05.

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Figure S4. Lactobacillus johnsonii (LJ) attenuated Escherichia coli (EC)-induced colitis and

memory impairment. BDNF expression and NF-κB activation was measured in the

hippocampus (a). Histological examination of colon tissues was performed by H&E and

immunostaining of MPO (b). For the histological examination, colons were fixed in 4%

paraformaldehyde, frozen, cut into 10-µm section using a cryostat, stained with

haematoxylin-eosin, and observed under a light microscopy. For the immunohistological

examination, the sections were detected using anti-CD86 antibody (a macrophage marker)

and 3,3’-diaminobenzene substrate kit. Horseradish peroxidase activity was visualized with

3-amino-9-ethylcarbazole. Colitis markers NF-κB activation, iNOS and COX-2 expression

(c), and tight junction protein expression (d), were measured in the colon. Proteins were

measured by immunoblotting. Values indicate mean± S.D. (n=10). *P<0.05.

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Figure S5. The difference of gut microbiota composition among normal control, TNBS-

treated, EC-treated, and EC plus LJ-treated groups. (A) The difference at the phylum level.

(b) The difference at the family level.

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Figure S7. Experimental design. Four animal experiments were performed. (A) The

preparation of mice with TNBS-induced colitis. Mice were intrarectally injected 2.5% (w/v)

TNBS solution (100 μL, dissolved in 50% ethanol). Normal control group was treated with

saline instead of TNBS. Mice were sacrificed 24 h, 8th day, and 15th day after TNBS

treatment. (B) The preparation of EC-induced colitis. Mice were orally administered EC (E.

coli) suspension (1 × 109 CFU, suspended in 100 μL of 1% glucose) once a day for 5 days.

Normal control group was treated with 1% glucose instead of EC. Mice were sacrificed 24 h,

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8th day, and 15th day after EC treatment. (C) The preparation of mice with LPS-induced

memory impairment. Mice were intraperitoneally injected LPS solution (8 μg/kg, dissolved

in saline) once a day for 10 days. LPS was isolated from EC. Normal control group was

treated with 1% glucose instead of EC. Mice were sacrificed 48 h after the final

administration of LPS treatment. (D) Evaluation for the anti-colitic and memory impairment-

ameliorating effect of LJ in mice with TNBS-induced colitis. Mice were randomly divided

into four groups: normal control, LJ-treated group in normal control, colitis control group

induced by treatment with TNBS, and LJ-treated group in mice with colitis. Normal control

group was treated with saline instead of TNBS. LJ (1 × 109 CFU/mouse, suspended in 0.1 mL

of 1% glucose) were orally administered once a day for 5 days from 72 h after the final

treatment with TNBS. Normal group was treated with 1% glucose (vehicle) alone instead of

LJ. (E) Evaluation for the anti-colitic and memory impairment-ameliorating effect of LJ in

mice with EC-induced colitis. Mice were randomly divided into four groups: normal control,

LJ-treated group in normal control, colitis control group induced by treatment with EC, and

LJ-treated group in mice with colitis. Normal control group was treated with saline instead of

EC. LJ (1 × 109 CFU/mouse, suspended in 0.1 mL of 1% glucose) were orally administered

once a day for 5 days from 72 h after the final treatment with EC. Normal group was treated

with 1% glucose (vehicle) alone instead of LJ. In all experiments, each group consisted of ten

mice. Memory behaviors were measured 2 h after the final administration of LJ in Y-maze

and passive avoidance task. Mice were killed 2 h after the measurement of memory

behaviors. The hippocampus and colon were removed. The colons were opened

longitudinally and gently washed with ice-cold PBS. The specimens were stored at -80 °C

until used in the experiment for the assays of enzyme activity, ELISA, and immunoblotting.

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