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: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: Proteins preferential to either HFpEF or control groups

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Control, Sequencing, Ubiquitin Proteomics

Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: Representative MS/MS scan for S100A8 peptide sequence ALNSIIDVYHK. Raw m/z spectral images with peak assignments and b and y ion lists along with a representation of peptide sequencing by tandem mass spectrometry

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Tandem Mass Spectroscopy, Sequencing, Mass Spectrometry

Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: Plasma levels of S100A8 in control vs. HFpEF groups. a S100A8 is found in increased levels in the plasma of subjects with HFpEF vs. control subjects as detected by ELISA. The MCW columns include the control (n = 7) and HFpEF (n = 9) from the discovery cohort and the NWU colums include the control (n = 18) and HFpEF (n = 25) samples from the validation cohort. *p < 0.006 vs MCW Control. # p < 0. 002 vs NWU Control

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Clinical Proteomics, Control, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: Overview of primary and secondary screening methods to identify potential mediators of HFpEF. a Platelet proteomes were subject to mass spectral analysis and novel proteins were identified. b Human cardiomyocytes derived from induced pluripotent stem cells were used to determine whether proteins that were identified in a had direct effects on cardiomyocytes function in vitro. Purified recombinant protein S100A8 was tested in this assay

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Derivative Assay, In Vitro, Purification, Recombinant

Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: S100A8-mediated effects on human iPSC-derived cardiomyocytes. a Shows example action potentials recorded from rS100A8 treated iPSC derived human cardiomyocytes. The addition of rS100A8 to the buffer extended the period between action potentials. This period is phase 4; the diastolic membrane potential between action potentials. b rS100A8 exacerbates the arrhythmic tendencies of human cardiomyocytes. c Spontaneous Ca 2+ transients recorded from human cardiomyocytes treated with rS100A8 as indicated by the blue line. rS100A8 significantly delayed the recovery of depolarization. Wash out of rS100A8 reversed these effects

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Derivative Assay, Membrane

Journal: Oncotarget

Article Title: Pancreatic cancer-derived exosomes promote tumor metastasis and liver pre-metastatic niche formation

doi: 10.18632/oncotarget.18831

Figure Lengend Snippet: Confocal microscopy (lung, liver, spleen, brain) of PKH67-labeled Panc02 EXO and Panc02-H7 EXO tissue distribution (green) 24 hpi. (A) PKH-67-labeled liposomes served as controls (scale bar=100 μm). Histogram shows exosome tissue distribution quantification (n=5/group). CD45, p-Stat3, and CD11b IF staining in liver sections from controls (left) and mice treated with Panc02 EXOs(middle) or Panc02-H7 EXOs (right) for 12 d without tumor challenge. (B) Histogram shows infiltrating CD45 + cell quantification. FN and α-SMA IF staining in liver sections from controls (top) and mice treated with Panc02 EXOs(middle) or Panc02-H7 EXOs (bottom) for 12 d without tumor challenge. (C) Histogram shows infiltrating α-SMA + hStCs and FN expression quantification(400× magnification; n=5/group). Western blotting analysis showed upregulated S100A8 and S100A9 in livers treated with Panc02-H7-derived exosomes. Histogram shows expression of the three proteins in three groupsas determined by densitometric analysis (n=3/group). (D) Pancreatic cancer-derived exosomes induce MDSC accumulation in peripheral blood. (E) Representative flow cytometric plots (left) and quantification (right) of CD11b + GR1 + MDSCs (n=5/group). *P<0.05, **P<0.01,***P<0.001.

Article Snippet: Primary antibodies against fibronectin (1:100),α-SMA (1:50), S100A8 (1:50), S100A9 (1:50), and F4/80 (1:50) were purchased from Proteintech (Wuhan, China).

Techniques: Confocal Microscopy, Labeling, Liposomes, Staining, Expressing, Western Blot, Derivative Assay

Journal: Oncotarget

Article Title: Pancreatic cancer-derived exosomes promote tumor metastasis and liver pre-metastatic niche formation

doi: 10.18632/oncotarget.18831

Figure Lengend Snippet: IHC analysis and histopathological examination of macrophages (F4/80), hStCs (α-SMA), and neutrophils in liver metastatic niches of naïve mice and mice treated with PBS, Panc02 EXOs, or Panc02-H7 EXOs at 30d post-SOI (arrow shows neutrophils in liver) (A) . Representative histogram shows quantification of F4/80 + macrophages, α-SMA + hStCs, and neutrophils (B) . Identification of FN, S100A8, and S100A9 as inflammatory mediators, and collagen deposition in the liver metastatic niche (C) . Representative histogram shows FN and MTS quantification (D) . Representative histogram shows S100A8 and S100A9 quantification (E) . n=6/group.**P<0.01,***P<0.001.10 fields assessed per sample. FOV, field of view.

Article Snippet: Primary antibodies against fibronectin (1:100),α-SMA (1:50), S100A8 (1:50), S100A9 (1:50), and F4/80 (1:50) were purchased from Proteintech (Wuhan, China).

Techniques:

Journal: Nature Communications

Article Title: Senescent immune cells accumulation promotes brown adipose tissue dysfunction during aging

doi: 10.1038/s41467-023-38842-6

Figure Lengend Snippet: a Bioinformatics analysis of scRNA-seq of BAT from young (5-month-old) and aged (27-month-old) rats. b GO analysis of differentially expressed genes in T cells (left) and neutrophils (right) from aged rats. c Violin plots for gene expression of S100a8 in cell populations of BAT from young and aged rats. d Relative cell proportion of S100A8 + and S100A8 − cell populations in the BAT of young and aged rats. e Violin plots for gene expression of S100a8 in T cell subclusters of young and aged rats. f UMAP plot shows clustering of macrophages based on gene expression. g Violin plots for gene expression of S100a8 in macrophage subclusters of young and old. h , i Representative flow cytometry plots and quantification of the frequencies of S100A8 + cells in CD45 + CD3 + T cells ( h ), and CD45 + CD11b + myeloid cells ( i ), in the stromal vascular fractions (SVFs) of BAT from 2-, 8- and 15-month-old mice ( n = 3–5/group). j Representative images of CD3 (green) and S100A8 (red) and staining in the BAT of young (2 months) and aged (15 months) mice. Scale bar, 50 µm. k Representative images of CD11b (green) and S100A8 (red) staining in the BAT of young (2 months) and aged (15 months) mice. Scale bar, 50 µm. Data shown are representative of three independent experiments with similar results. n indicates the number of biologically independent samples examined. Quantitative data are shown as mean ± SEM. Statistical differences were supposed to be significant when P < 0.05. Statistical analysis was performed by one-way ANOVA with Tukey’s multiple-comparison test ( h , i ). Source data are provided as a Source Data File.

Article Snippet: For recombinant S100A8 protein treatment, S100A8 protein (SinoBiological, 50228-M07E) was dissolved in PBS and administrated by tail vein injection at the dose of 2.5 μg per mouse twice a week for 4 weeks.

Techniques: Expressing, Flow Cytometry, Staining

Journal: Nature Communications

Article Title: Senescent immune cells accumulation promotes brown adipose tissue dysfunction during aging

doi: 10.1038/s41467-023-38842-6

Figure Lengend Snippet: a Schematic diagram for isolation, labeling and transferring bone marrow-derived S100A8 + immune cells into young mice. This diagram was created with BioRender.com. b Representative images of PKH26-labeled S100A8 − or S100A8 + immune cells infiltrating BAT, iWAT, eWAT and liver ( n = 3). Scale bar, 100 μm. c Schematic diagram for bone marrow reconstitution of mice with HSPCs isolated from S100a8-Cre-EGFP mice. This diagram was created with BioRender.com. d Gating strategy of bone marrow hematopoietic stem/progenitor cells (HSPCs). e Representative images of GFP + cells in the BAT of bone marrow reconstitution mice during aging ( n = 4). Scale bar, 100 μm. f , g Representative flow cytometry plots and quantification of the frequencies of GFP + cells in CD45 + CD3 + T cells ( f ) ( n = 5/group), and CD45 + CD11b + myeloid cells ( g ) ( n = 5/group), in the SVFs of BAT from bone marrow reconstitution mice during aging. Data shown are representative of three independent experiments with similar results. n indicates the number of biologically independent samples examined. Data are shown as mean ± SEM. Statistical differences were supposed to be significant when P < 0.05. Statistical analysis was performed by one-way ANOVA with Tukey’s multiple-comparison test ( e – g ). Source data are provided as a Source Data File.

Article Snippet: For recombinant S100A8 protein treatment, S100A8 protein (SinoBiological, 50228-M07E) was dissolved in PBS and administrated by tail vein injection at the dose of 2.5 μg per mouse twice a week for 4 weeks.

Techniques: Isolation, Labeling, Transferring, Derivative Assay, Flow Cytometry

Journal: Nature Communications

Article Title: Senescent immune cells accumulation promotes brown adipose tissue dysfunction during aging

doi: 10.1038/s41467-023-38842-6

Figure Lengend Snippet: a Representative immunoblots of thermogenic markers UCP1 and PGC-1α in the BAT of mice transferred with S100A8 + immune cells or S100A8 − immune cells ( n = 4). b Representative images of UCP1 staining of the BAT from mice transferred with S100A8 + immune cells or S100A8 − immune cells. Scale bar, 50 μm. c Relative mRNA levels of Ucp1 , Ppargc1α, p16 and p21 in the BAT of mice transferred with S100A8 + immune cells or S100A8 − immune cells ( n = 4). d Representative immunoblots of P16 and P21 in the BAT of mice transferred with S100A8 + immune cells or S100A8 − immune cells ( n = 5). e Oxygen consumption of mice transferred with S100A8 + immune cells or S100A8 − immune cells ( n = 6–9). f Energy expenditure of mice transferred with S100A8 + immune cells or S100A8 − immune cells ( n = 6–9). g Core body temperature of mice transferred with S100A8 + immune cells or S100A8 − immune cells under cold stimulation ( n = 5). h Hematoxylin-eosin staining and adipocyte cell-diameter quantification for BAT of mice transferred with S100A8 + immune cells or S100A8 - immune cells. Scale bar, 50 μm. i Relative mRNA levels of thermogenic and aging-related genes in the BAT of mice injected with AAV-Scramble and AAV-Sh S100a8 ( n = 5). j Core body temperature of mice injected with AAV-Scramble and AAV- Sh S100a8 under cold stimulation ( n = 5). Data shown are representative of three independent experiments with similar results. n indicates the number of biologically independent samples examined. Data are shown as the mean ± SEM. Statistical differences were supposed to be significant when P < 0.05. Statistical analysis was performed by two-way ANOVA ( g , j ), ANCOVA with body weight as covariant ( e , f ) or unpaired two-tailed Student’s t test ( a , c , d , i ). Source data are provided as a Source Data File.

Article Snippet: For recombinant S100A8 protein treatment, S100A8 protein (SinoBiological, 50228-M07E) was dissolved in PBS and administrated by tail vein injection at the dose of 2.5 μg per mouse twice a week for 4 weeks.

Techniques: Western Blot, Staining, Injection, Two Tailed Test

Journal: Nature Communications

Article Title: Senescent immune cells accumulation promotes brown adipose tissue dysfunction during aging

doi: 10.1038/s41467-023-38842-6

Figure Lengend Snippet: a Heatmap of differentially expressed genes (DEGs) in differentiated brown adipocytes treated with S100A8 ( P < 0.05 and |log 2 (Fold change)| >0.38). b GO analysis of DEGs in ( a ). c Representative images of age-associated changes in S100A8 + immune cells and tyrosine hydroxylase positive (TH + ) sympathetic nerves in the BAT of 2-month-old and 15-month-old mice. Scale bar, 100 μm. d Representative images of the localization of S100A8 + immune cells and TH + sympathetic nerves in the BAT of 2- and 15-month-old mice. Scale bar, 10 μm. e Pearson correlation analysis of the fluorescence intensity between S100A8 and TH in the BAT of 2-month-old and 15-month-old mice. f Representative immunoblots of TH, phospho-TH (Ser40) (p-TH) and TUBB3 in the BAT of mice transferred with S100A8+ immune cells or S100A8- immune cells ( n = 3). g Schematic diagram of neuroimmune adipose interface in the BAT. This diagram was created with BioRender.com. h Venn diagram of downregulated DEGs in RNA-seq datasets of S100A8-treated brown adipocytes and BAT of aged mice. i Representative immunoblots of RBM3 in the BAT of 2- and 15-month-old mice ( n = 3). j Representative immunoblots of RBM3 in the BAT of mice transferred with S100A8 + immune cells or S100A8 − immune cells ( n = 3). k Representative images of TH staining in the BAT of mice injected with AAV-Sh Rbm3 or AAV-Scramble. Scale bar, 100 µm. l Representative immunoblots of UCP1, TH, p-TH, TUBB3 and RBM3 in the BAT of mice injected with AAV-Sh Rbm3 or AAV-Scramble ( n = 3). m Core body temperature of mice injected with AAV-Sh Rbm3 or AAV-Scramble under cold stimulation ( n = 5–7). Data shown are representative of three independent experiments with similar results. n indicates the number of biologically independent samples examined. Data are shown as the mean ± SEM. Statistical differences were supposed to be significant when P < 0.05. Statistical analysis was performed by two-way ANOVA ( m ), or unpaired two-tailed Student’s t test ( f , i – l ). Source data are provided as a Source Data File.

Article Snippet: For recombinant S100A8 protein treatment, S100A8 protein (SinoBiological, 50228-M07E) was dissolved in PBS and administrated by tail vein injection at the dose of 2.5 μg per mouse twice a week for 4 weeks.

Techniques: Fluorescence, Western Blot, RNA Sequencing Assay, Staining, Injection, Two Tailed Test

Journal: Nature Communications

Article Title: Senescent immune cells accumulation promotes brown adipose tissue dysfunction during aging

doi: 10.1038/s41467-023-38842-6

Figure Lengend Snippet: a Schematic diagram of experimental processes. This diagram was created with BioRender.com. b Representative images of TH staining in the BAT of mice injected with AAV-RBM3 or AAV-NC and transferred with S100A8 + immune cells. Scale bar, 100 µm. c Representative immunoblots of UCP1, TH, p-TH, TUBB3 and RBM3 in the BAT of mice injected with AAV-RBM3 or AAV-NC and transferred with S100A8 + immune cells ( n = 3). d Relative mRNA levels of Rbm3, Th Ucp1, Ppargc1α, p16, and p21 in the BAT of mice injected with AAV-RBM3 or AAV-NC and transferred with S100A8 + immune cells ( n = 5). e Oxygen consumption of mice injected with AAV-RBM3 or AAV-NC and transferred with S100A8 + immune cells or S100A8 - immune cells ( n = 6–7). f Core body temperature of mice injected with AAV-RBM3 or AAV-NC were transferred with S100A8 + immune cells and subjected to cold challenge ( n = 5). g Venn diagram showing overlapped genes which are RBM3-bounded and differentially expressed upon Rbm3 knockdown. h KEGG analysis of the 909 overlapped genes shown in ( g ). i Heatmap of axon guidance-related gene expressions based on RNA-seq data of Rbm3 knockdown differentiated brown adipocytes. j Relative mRNA levels of Rbm3 and axon guidance-related genes Nrp1 and Epha7 in differentiated brown adipocytes transfected with si- Rbm3 or si-NC ( n = 3). k Representative immunoblots of NRP1 and EPHA7 in differentiated brown adipocytes transfected with si- Rbm3 or si-NC ( n = 4). l RNA immunoprecipitation (RIP) assay assessing RBM3 binding on 3´UTR of Nrp1 and Epha7 in differentiated brown adipocytes ( n = 4). m Relative mRNA level of Nrp1 and Epha7 in si-Rbm3 or si-NC transfected brown adipocytes upon transcriptional inhibition with actinomycin D at indicated time ( n = 3). n Representative images of TUBB3 staining in PC12 cells cocultured with differentiated brown adipocytes overexpressed with RBM3 and transfected with si- Nrp1 or si- Epha7 ( n = 4). Scale bar, 100 µm. Data shown are representative of three independent experiments with similar results. n indicates the number of biologically independent samples examined. Data are shown as the mean ± SEM. Statistical differences were supposed to be significant when P < 0.05. Statistical analysis was performed by one-way ANOVA with Tukey’s multiple-comparison test ( n ) or two-way ANOVA ( f , m ), ANCOVA with body weight as covariant ( e ) or unpaired two-tailed Student’s t test ( c , d , j – l ). Source data are provided as a Source Data File.

Article Snippet: For recombinant S100A8 protein treatment, S100A8 protein (SinoBiological, 50228-M07E) was dissolved in PBS and administrated by tail vein injection at the dose of 2.5 μg per mouse twice a week for 4 weeks.

Techniques: Staining, Injection, Western Blot, RNA Sequencing Assay, Transfection, Immunoprecipitation, Binding Assay, Inhibition, Two Tailed Test

Journal: Nature Communications

Article Title: Senescent immune cells accumulation promotes brown adipose tissue dysfunction during aging

doi: 10.1038/s41467-023-38842-6

Figure Lengend Snippet: a , b Representative flow cytometry plots and quantification of frequencies of S100A8 + immune cells ( a ) and T cells ( b ) in peripheral blood cells from young and old normal healthy donors (all males; Young: n = 9, Old: n = 13). c Correlation of S100a8 with p21 ( Kras ) expression in human whole blood cells based on data from the Genotype Tissue Expression (GTEx) database. d Schematic diagram of experimental processes of transplantation of human S100A8 + or S100A8 − immune cells into NOD-SCID mice. This diagram was created with BioRender.com. e Serum concentration of TNF and IL-6 in transplanted mice ( n = 5). f Representative flow cytometry plots and quantification of frequencies of human CD45 + immune cells in the SVFs of BAT from transplanted mice ( n = 5). g Representative images of TH and human CD45 staining in the BAT of transplanted mice. Scale bar, 100 μm. h Representative immunoblots of UCP1, P16, and P21 in the BAT of transplanted mice ( n = 5). i SA-β-gal staining of BAT and its frozen sections of transplanted mice. j Representative infrared thermal images of BAT of transplanted mice subjected to cold stimulation for 6 h ( n = 5). k Core body temperature of transplanted mice under cold stimulation ( n = 5). Data shown are representative of three independent experiments with similar results. n indicates the number of biologically independent samples examined. Data are shown as the mean ± SEM. Statistical differences were supposed to be significant when P < 0.05. Statistical analysis was performed by two-way ANOVA ( k ), or unpaired two-tailed Student’s t test ( a , b , e – h ). Source data are provided as a Source Data File.

Article Snippet: For recombinant S100A8 protein treatment, S100A8 protein (SinoBiological, 50228-M07E) was dissolved in PBS and administrated by tail vein injection at the dose of 2.5 μg per mouse twice a week for 4 weeks.

Techniques: Flow Cytometry, Expressing, Transplantation Assay, Concentration Assay, Staining, Western Blot, Two Tailed Test

Journal: Nature Communications

Article Title: Senescent immune cells accumulation promotes brown adipose tissue dysfunction during aging

doi: 10.1038/s41467-023-38842-6

Figure Lengend Snippet: a , b Representative flow cytometry plots and quantification of frequencies of S100A8 + T cells ( a ) and myeloid cells ( b ) in SVFs of BAT of 15-month-old mice treated with paquinimod or vehicle ( n = 4–5). c Representative images of TH and S100A8 staining in the BAT of 15-month-old mice treated with paquinimod or vehicle. Scale bar, 20 µm. d Relative mRNA levels of Th and Ucp1 in the BAT of 15-month-old mice treated with paquinimod or vehicle ( n = 5). e Representative immunoblots of UCP1 and PGC-1α in the BAT of 15-month-old mice treated with paquinimod or vehicle ( n = 6). f Representative immunoblots of P16 and P21 in the BAT of 15-month-old mice treated with paquinimod or vehicle ( n = 3). g Food intake of mice treated with paquinimod or vehicle ( n = 5). h Body weight changes of normal chow diet-fed 15-month-old mice treated with paquinimod or vehicle ( n = 5–6). i Fasting glucose levels of normal chow diet-fed mice after treated with paquinimod or vehicle for 6 months ( n = 5–6). j – n Body weight changes ( j ), glucose tolerance ( k ), insulin sensitivity ( l ), tissue weights ratio ( m ), HE staining of adipose tissue and liver ( n ) in HFD-fed mice treated with paquinimod or vehicle ( n = 5). Scale bar, 50 µm. Data shown are representative of three independent experiments with similar results. n indicates the number of biologically independent samples examined. Data are shown as the mean ± SEM. Statistical differences were supposed to be significant when P < 0.05. Statistical analysis was performed by two-way ANOVA ( h , j – l ), or unpaired two-tailed Student’s t test ( a , b , d – f , i , m ). Source data are provided as a Source Data File.

Article Snippet: For recombinant S100A8 protein treatment, S100A8 protein (SinoBiological, 50228-M07E) was dissolved in PBS and administrated by tail vein injection at the dose of 2.5 μg per mouse twice a week for 4 weeks.

Techniques: Flow Cytometry, Staining, Western Blot, Two Tailed Test