Fig 2: Effect of exendin-4 treatment on the expression of inflammatory cytokines by mouse CSMCs using antibody array membrane. (A) Freshly isolated CSMCs from 20 male Balb/C mice were cultured in DMEM and divided into four treatment groups: control, LPS, LPS+exendin-4 (50 nM), and LPS+exendin-4 (100 nM). After 24 h of incubation the cells were collected, lysed, and incubated with the antibody array membrane. There was a basal expression of various cytokines by the normal CSMCs which was increased once inflammation was induced by LPS. Exendin-4 at both concentrations inhibited the overproduction of cytokines induced by inflammation. A visual comparison between the four arrays performed by two independent investigators and by ImageJ software (National Institutes of Health) revealed that five cytokines were especially affected by the treatment: tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), T cell activation gene-3 (TCA-3), stromal cell-derived factor-1 (SDF-1), and macrophage colony stimulating factor (M-CSF). (B) List of cytokines and chemokines represented by the antibody array membrane (Cat. No. ab133999; Abcam Inc.).Abbreviations: CSMC, colon smooth muscle cell; Ex 50, exendin-4 (50 nM); Ex 100, exendin-4 (100 nM); GCSF, granulocyte colony stimulating factor; GM-CSF, granulocyte– macrophage colony stimulating factor; IFN, interferon; IL, interleukin; LPS, lipopolysaccharide; Pos, positive; Neg, negative.

Fig 3: Effect of exendin-4 treatment on the expression of inflammatory cytokines by mouse CSMCs using antibody array membrane. (A) Freshly isolated CSMCs from 20 male Balb/C mice were cultured in DMEM and divided into four treatment groups: control, LPS, LPS+exendin-4 (50 nM), and LPS+exendin-4 (100 nM). After 24 h of incubation the cells were collected, lysed, and incubated with the antibody array membrane. There was a basal expression of various cytokines by the normal CSMCs which was increased once inflammation was induced by LPS. Exendin-4 at both concentrations inhibited the overproduction of cytokines induced by inflammation. A visual comparison between the four arrays performed by two independent investigators and by ImageJ software (National Institutes of Health) revealed that five cytokines were especially affected by the treatment: tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), T cell activation gene-3 (TCA-3), stromal cell-derived factor-1 (SDF-1), and macrophage colony stimulating factor (M-CSF). (B) List of cytokines and chemokines represented by the antibody array membrane (Cat. No. ab133999; Abcam Inc.).Abbreviations: CSMC, colon smooth muscle cell; Ex 50, exendin-4 (50 nM); Ex 100, exendin-4 (100 nM); GCSF, granulocyte colony stimulating factor; GM-CSF, granulocyte– macrophage colony stimulating factor; IFN, interferon; IL, interleukin; LPS, lipopolysaccharide; Pos, positive; Neg, negative.

Fig 4: Effect of exendin-4 treatment on the expression of inflammatory cytokines by mouse CSMCs using antibody array membrane. (A) Freshly isolated CSMCs from 20 male Balb/C mice were cultured in DMEM and divided into four treatment groups: control, LPS, LPS+exendin-4 (50 nM), and LPS+exendin-4 (100 nM). After 24 h of incubation the cells were collected, lysed, and incubated with the antibody array membrane. There was a basal expression of various cytokines by the normal CSMCs which was increased once inflammation was induced by LPS. Exendin-4 at both concentrations inhibited the overproduction of cytokines induced by inflammation. A visual comparison between the four arrays performed by two independent investigators and by ImageJ software (National Institutes of Health) revealed that five cytokines were especially affected by the treatment: tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), T cell activation gene-3 (TCA-3), stromal cell-derived factor-1 (SDF-1), and macrophage colony stimulating factor (M-CSF). (B) List of cytokines and chemokines represented by the antibody array membrane (Cat. No. ab133999; Abcam Inc.).Abbreviations: CSMC, colon smooth muscle cell; Ex 50, exendin-4 (50 nM); Ex 100, exendin-4 (100 nM); GCSF, granulocyte colony stimulating factor; GM-CSF, granulocyte– macrophage colony stimulating factor; IFN, interferon; IL, interleukin; LPS, lipopolysaccharide; Pos, positive; Neg, negative.

Fig 5: Effect of exendin-4 treatment on the expression of inflammatory cytokines by mouse CSMCs using antibody array membrane. (A) Freshly isolated CSMCs from 20 male Balb/C mice were cultured in DMEM and divided into four treatment groups: control, LPS, LPS+exendin-4 (50 nM), and LPS+exendin-4 (100 nM). After 24 h of incubation the cells were collected, lysed, and incubated with the antibody array membrane. There was a basal expression of various cytokines by the normal CSMCs which was increased once inflammation was induced by LPS. Exendin-4 at both concentrations inhibited the overproduction of cytokines induced by inflammation. A visual comparison between the four arrays performed by two independent investigators and by ImageJ software (National Institutes of Health) revealed that five cytokines were especially affected by the treatment: tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), T cell activation gene-3 (TCA-3), stromal cell-derived factor-1 (SDF-1), and macrophage colony stimulating factor (M-CSF). (B) List of cytokines and chemokines represented by the antibody array membrane (Cat. No. ab133999; Abcam Inc.).Abbreviations: CSMC, colon smooth muscle cell; Ex 50, exendin-4 (50 nM); Ex 100, exendin-4 (100 nM); GCSF, granulocyte colony stimulating factor; GM-CSF, granulocyte– macrophage colony stimulating factor; IFN, interferon; IL, interleukin; LPS, lipopolysaccharide; Pos, positive; Neg, negative.