Presentación de PowerPoint - Nature Research0,2 0,4 0,6 0,8 1 WT KO WT+DSS KO+DSS y *** Figure S3. KI67 staining (in red; and nuclei in blue) in untreated and DSS-treated WT and cortactin

ctrl DSS

WT

KO

Supplementary information

Figure S1. Histological scans and scores of whole colon “swiss roles”.

(A) Representative images from the 4 experimental groups after five days are shown (Bars=2mm).

(B) Histological scores of the 4 experimental groups determined in a blinded fashion as described

in materials and methods.

A B

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Figure S2. Goblet cell staining and quantification.

(A) Periodic acid-Schiff stainings of paraffin-embedded colon tissues. Representative images of distal

colon areas from 3 independent preparations are shown. Bar = 50 µm.

(B) Quantification of goblet cell numbers. Goblet cells were counted in random 500 µm areas of distal

colons. Four of such areas were analyzed per swiss rolls from 3 different mice in each group.

(C) Circularity of goblet cells (a score of 1 represents round cells). 50 random goblet cells of distal

colons were measured per slide in 3 independent preparations using the circularity tool of Image J.

Data are displayed as means+/-SDM. *p<0.05; **p<0.01; ***p<0.001.

B C

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Figure S3. KI67 staining (in red; and nuclei in blue) in untreated and DSS-treated WT and

cortactin KO mice 5 days after starting the experiment. Under inflammatory conditions,

cortactin deficiency no longer causes increased proliferation. Images representative of

3 independent experiments. Bar = 50 µm.

WT KO WT+DSS KO+DSS

Figure S4. Immunohistochemical stainings for leukocytes using antibodies against CD45 and CD68.

(A, B) Representative images of CD45 (A) and CD68 (B) stainings of the 4 experimental groups (n=5)

after five days of starting the experiment are shown. Positive leukocytes appear dark brown.

Nuclei are shown in blue. Bars=50 µm.

(C,D) Quantification of CD45-positive (C) and CD68-positive (D) leukocytes in the colons of the 4

experimental groups. Leukocytes were counted in 10 random images taken of the distal colon

of 3 independent stainings. Data are displayed as mean leukocyte numbers per image+/-SDM.

*p<0.05; **p<0.01; ***p<0.001.

A

B

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Figure S5. Brightfield images of cortactin-depleted (KD) and scrambled (scr) control Caco-2

cells taken every 24 h after plating 1,8x105 cells in each well of a 24-well plate. Images are

representative of 3 independent experiments. Bar = 100 µm.

scr

KD

24 h 48 h 72 h

Figure S6. Representative images (n=3) of cortactin-depleted and scrambled (scr) control Caco-2

stained for actin (red) and ZO-1 (green). Images show a clear increase in actin fibers in the apical

regions of cortactin-depleted cells. Actin and ZO-1 colocalization appears yellow. Bar = 20 µm.

actin ZO-1 merge actin ZO-1 merge

shRNA scr shRNA cortactin

total view

apical

basal

center

Figure S7. Representative images (n=3) of cortactin KO and WT colon tissue cross-sections stained for

actin (red) and ZO-1 (green). Images show a clear increase in actin fibers and internalization of ZO-1.

Bar = 50 µm.

WT

KO

actin ZO-1 nuclei merge