Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of estrogen treatment on S100A8 expression in endometrium and epithelium morphology. ( a ) S100A8 immunohistochemical staining before and after estrogen treatment (1d/7d); black arrows indicate S100A8-positive immune cells (S100A8-PICs).The blue arrow indicates the release of S100A8 from S100A8-PICs. Scale bar is 100 or 400 μm. ( b ) Rat endometrial hematoxylin–eosin staining before and after estrogen treatment (1d/7d); red arrows show disrupted cell junctions. Scale bar is 100 μm. ( c ) S100A8-PICs number in the endometrium (1/mm 2 ) before and 1 d/7d after estrogen treatment (n = 18, from six rats, the number was counted from three immunohistochemical images of each sample , mean ± SEM). ( d ) S100A8 expression in the epithelium before and 1 d/7d after estrogen treatment (n = 18, from six rats, the expression level was measured from three immunohistochemical images of each sample, mean ± SEM). * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of uterine cavity administration of S100A8 on the distribution of S100A8-PICs and endometrial morphology. ( a ) Distribution of S100A8-positive cells in the endometrium of four different groups with estrogen treatment (control, injury, P407, and S100A8 + P407 groups). Black arrows show S100A8-PICs. S100A8-PICs were distributed around the glands and blood vessels in the control group. S100A8-PICs in the site of injury or adhesion in the injury group and in the P407 group. S100A8-PICs reverse migrated to the glands or blood vessels in the S100A8 + P407 group. Scale bar is 100 μm or 1 mm. ( b ) Endometrial thickness in the four groups (n = 15, from 5 rats; 3 position was measured randomly in an image of each rat, mean ± SEM). ( c ) The number of endometrial glands in each group (n = 10, from 10 rats, glands were counted from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). ( d ) The interior wall integrity of uterine cavity in each group (n = 10, from 10 rats, data was measured from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). ( e ) The number of endometrial crypts in each group (n = 10, from 10 rats, crypts were counted from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Control

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on the distribution of Ki-67-positive proliferating cells in the uteri and the proliferation of endometrial cells. ( a ) Ki-67-positive proliferating cells were distributed in the endometrium or uterine interior wall in all groups (control, injury, P407, and S100A8 + P407 groups). Scale bar is 100 or 400 μm. ( b ) Number of Ki-67-positive cells in the endometrium of each group (n = 20, from ten rats, the number was counted from two immunohistochemical images of each sample, mean ± SEM). ( c ) Number of Ki-67-positive cells in the uterine interior wall of each group (n = 20, from ten rats, the number was counted from two immunohistochemical images of each sample, mean ± SEM). ( d ) Effects of S100A8 and its receptor inhibitor (FPS-ZM1) on the proliferation of cultured endometrial cells in vitro (n = 5, mean ± SEM). Scale bar is 100 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Control, Immunohistochemical staining, Cell Culture, In Vitro

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on tight junctions of endometrial epithelium or endometrial cells. ( a ) Immunohistochemical staining showing Claudin-1 distribution in the endometrium of different groups (control, injury, P407, and P407 + S100A8 groups); black arrows show Claudin-1 accumulated below the cell membrane of the lateral contact zone between adjacent cells and red arrows show cells expressing Claudin-1 in the stroma. Scale bar is 100 μm. ( b ) Claudin-1 expression in the endometrium of different groups (n = 10, mean ± SEM). ( c ) Western blot showing ZO-1 expression in endometrial cells of three different groups (control, S100A8, and S100A8 + FPS-ZM1 groups, n = 3, mean ± SEM). Actin was used as the loading control. The full-length bands can be found in supplementary Fig. 1. The bands of ZO-1 and actin cropped from different parts of the same gel. ( d ) Immunofluorescence showing the co-expression of Claudin-1 (red) and ZO-1 (green) in the endometrial cells of the three groups (n = 5, mean ± SEM). The nucleus is stained blue. The scale bar is 200 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Immunohistochemical staining, Staining, Control, Membrane, Expressing, Western Blot, Immunofluorescence

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on endometrial “double-featured” cells. ( a ) Vimentin (red) and ZO-1 (green) immunofluorescent double staining of cultured endometrial cells. Scale bar is 200 μm. ( b ) Western blot showing CK-18 and vimentin expression in endometrial cells from each group (control, S100A8, and FPS-ZM1 groups; n = 3, mean ± SEM). The full-length bands can be found in supplementary Fig. 1. Actin was used as the loading control. As the target proteins have a similar molecular weight to that of the loading control, the bands of CK18/vimentin and actin were cropped from different gels. ( c ) Immunofluorescence showing vimentin expression in endometrial cells of each group (n = 5, mean ± SEM). Scale bar is 200 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Double Staining, Cell Culture, Western Blot, Expressing, Control, Molecular Weight, Immunofluorescence

Journal: Acta dermato-venereologica

Article Title: Crisaborole Inhibits Itch and Pain by Preventing Neutrophil Infiltration in a Mouse Model of Atopic Dermatitis.

doi: 10.2340/actadv.v103.13382

Figure Lengend Snippet: Fig. 2. Effects of crisaborole on neutrophil infiltration in atopic dermatitis mouse model. (A–F) Skin was dissected from mice on day 8 after EtOH (A, E, F), MC903 (MC: B, E, F), MC903+vehicle (MC+VH: C, E, F), or MC903+crisaborole treatment (MC+Cris: D, E, F). Skin sections were immunostained with an antibody for PGP9.5 and Ly6G to visualize nerve fibres (green) and neutrophils (red), respectively. Dotted lines indicate the dermal-epidermal junction. White arrows indicate neutrophils contact with nerve fibres. (E) Scale bar indicates 20 µm. Total number of neutrophils in the epidermis was quantified in EtOH (white bars), MC (green bars), MC+VH (dark grey bars), and MC+Cris mice (blue bars). Error bars are SEM. **p < 0.01, ***p < 0.001, ****p < 0.0001, significant difference (1-way analysis of variance (ANOVA) followed by Tukey test, n = 5/group). (F) As in (E) for epidermal neutrophils in contact with nerves. (G and H) Skin sections were immunostained with an antibody for Ly6G and S100A8 or S100A9.

Article Snippet: Skin sections were incubated with 5% goat or donkey serum and 0.2% Triton X-100 in PBS, and then immunostained with a primary antibody directed against PGP 9.5 (1:2000; Chemicon (Millipore), Billerica, MA, USA; Cat# AB1761-I, RRID:AB_2868444), to label all epidermal nerve fibres at 4°C overnight, followed by incubation with the corresponding secondary antibody conjugated with AlexaFluor 488 (1:300; Life Technologies Inc., Grand Island, NY, USA) for 2 h. Subsequently, the sections were immunostained with rat Ly6G antibody (1:500; Bio X Cell, Lebanon, NH, USA; Cat# BE0075-1, RRID:AB_1107721), the most commonly used marker for neutrophils at 4°C overnight, followed by incubation with the corresponding secondary antibody conjugated with AlexaFluor 633 (1:300; Life Technologies Inc.) for 2 h. For S100A8 or S100A9 experiments, the sections were incubated with goat S100A9 antibody (1:400; R&D systems, Minneapolis, MN, USA; Cat# AF2065, RRID:AB_2184263) or rat S100A8 antibody (1:50; R&D Systems; Cat# MAB3059, RRID:AB_2184252) followed by goat secondary antibody conjugated with AlexaFluor 555 (1:300; Life Technologies Inc.) or rat secondary antibody conjugated with AlexaFluor 633 (1:300; Life Technologies Inc.) for 2 h, respectively.

Techniques:

Journal: Acta dermato-venereologica

Article Title: Crisaborole Inhibits Itch and Pain by Preventing Neutrophil Infiltration in a Mouse Model of Atopic Dermatitis.

doi: 10.2340/actadv.v103.13382

Figure Lengend Snippet: Fig. 3. S100A8/A9 facilitates pruritogen- or algogen-evoked scratching and wiping behaviours in mice. (A) S100A8/A9 (1, 10, 100 ng) or vehicle (phosphate-buffered saline; PBS) was intradermally injected into the nape of the neck. Following the injection, scratch bouts were counted over a 30-min period. Error bars are standard error of mean (SEM) n = 6–8. (B, C) S100A8/A9 (1, 10 ng) or vehicle was intradermally injected into the nape of the neck with either histamine (54 nmol: B) or chloroquine (97 nmol: C). Error bars are SEM n = 6–8. *p < 0.05, **p < 0.01, ****p < 0.0001, significant difference from the vehicle-treated group. One-way analysis of variance (ANOVA) followed by the post hoc Tukey test. F (2, 19)=18.60, p < 0.0001 for histamine. F (2, 15)=6.353, p = 0.01 for chloroquine. (D–I) S100A8/A9 (10 ng) or vehicle was intradermally injected into the cheek with either histamine (54 nmol: D, G), chloroquine (97 nmol: E, H), or capsaicin (33 nmol: F, I). Error bars are SEM n = 5–6. *p < 0.05, ***p < 0.001, ****p < 0.0001, significant difference from the vehicle-treated group. Two-tailed unpaired t-test.

Article Snippet: Skin sections were incubated with 5% goat or donkey serum and 0.2% Triton X-100 in PBS, and then immunostained with a primary antibody directed against PGP 9.5 (1:2000; Chemicon (Millipore), Billerica, MA, USA; Cat# AB1761-I, RRID:AB_2868444), to label all epidermal nerve fibres at 4°C overnight, followed by incubation with the corresponding secondary antibody conjugated with AlexaFluor 488 (1:300; Life Technologies Inc., Grand Island, NY, USA) for 2 h. Subsequently, the sections were immunostained with rat Ly6G antibody (1:500; Bio X Cell, Lebanon, NH, USA; Cat# BE0075-1, RRID:AB_1107721), the most commonly used marker for neutrophils at 4°C overnight, followed by incubation with the corresponding secondary antibody conjugated with AlexaFluor 633 (1:300; Life Technologies Inc.) for 2 h. For S100A8 or S100A9 experiments, the sections were incubated with goat S100A9 antibody (1:400; R&D systems, Minneapolis, MN, USA; Cat# AF2065, RRID:AB_2184263) or rat S100A8 antibody (1:50; R&D Systems; Cat# MAB3059, RRID:AB_2184252) followed by goat secondary antibody conjugated with AlexaFluor 555 (1:300; Life Technologies Inc.) or rat secondary antibody conjugated with AlexaFluor 633 (1:300; Life Technologies Inc.) for 2 h, respectively.

Techniques: Saline, Injection, Two Tailed Test

Journal: Acta dermato-venereologica

Article Title: Crisaborole Inhibits Itch and Pain by Preventing Neutrophil Infiltration in a Mouse Model of Atopic Dermatitis.

doi: 10.2340/actadv.v103.13382

Figure Lengend Snippet: Fig. 4. Effects of S100A8/A9 on pruritogen- or algogen-induced calcium response in mouse dorsal root ganglion (DRG) neurones. (A) Time-course graph of histamine-induced calcium response in DRG neurones. DRG neurones were treated with vehicle (black line) or S100A8/A9 (blue line) with a subsequent challenge with histamine (100 μM). (B) As in (A) for chloroquine (100 μM). (C) As in (A) for capsaicin (1 μM). (D) Averaged area under the curve of the calcium responses to histamine. DRG neurones were treated with vehicle (black column) or S100A8/A9 (blue column). Error bars are SEM n = 19–27. ***p < 0.001, a significant difference from the vehicle group. 2-tailed unpaired t-test. (E) As in (D) for chloroquine. n = 11–22. (F) As in (D) for capsaicin. n = 13–14. (G) Proportions of histamine-, chloroquine-, or capsaicin-responsive DRG neurones pre-treated with vehicle (black column) or S100A8/A9 (blue column). *p < 0.05. Fisher exact test. n = 69–128.

Article Snippet: Skin sections were incubated with 5% goat or donkey serum and 0.2% Triton X-100 in PBS, and then immunostained with a primary antibody directed against PGP 9.5 (1:2000; Chemicon (Millipore), Billerica, MA, USA; Cat# AB1761-I, RRID:AB_2868444), to label all epidermal nerve fibres at 4°C overnight, followed by incubation with the corresponding secondary antibody conjugated with AlexaFluor 488 (1:300; Life Technologies Inc., Grand Island, NY, USA) for 2 h. Subsequently, the sections were immunostained with rat Ly6G antibody (1:500; Bio X Cell, Lebanon, NH, USA; Cat# BE0075-1, RRID:AB_1107721), the most commonly used marker for neutrophils at 4°C overnight, followed by incubation with the corresponding secondary antibody conjugated with AlexaFluor 633 (1:300; Life Technologies Inc.) for 2 h. For S100A8 or S100A9 experiments, the sections were incubated with goat S100A9 antibody (1:400; R&D systems, Minneapolis, MN, USA; Cat# AF2065, RRID:AB_2184263) or rat S100A8 antibody (1:50; R&D Systems; Cat# MAB3059, RRID:AB_2184252) followed by goat secondary antibody conjugated with AlexaFluor 555 (1:300; Life Technologies Inc.) or rat secondary antibody conjugated with AlexaFluor 633 (1:300; Life Technologies Inc.) for 2 h, respectively.

Techniques: