anti stip1 homology  (Proteintech)

Journal: Communications Biology

Article Title: Stress-inducible phosphoprotein 1 (STIP1) is a critical stemness regulator in mouse embryonic stem cells and early mammalian development

doi: 10.1038/s42003-025-08763-9

Figure Lengend Snippet: A Cells from the zygote to the late blastocyst projected onto a principal component analysis (PCA) plot and colored according to their developmental stage. B Violin plots with boxplot display Stip1 expression (normalized to zygote levels) across pre-implantation stages. Wilcoxon test, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001 for specific stages relative to all others. Boxplots depict the quartiles and extend from the minimum and maximum data points within 1.5 times the interquartile range. C PCA shown in ( A ), colored to indicate the scaled expression of Stip1 , Sox2, Pou5f1/Oct4 , Nanog and Klf4 . D Pearson correlation plot highlighting genes that are positively (red) or negatively (blue) correlated with Stip1 , while non-significant correlations are shown in light gray. E Gene Ontology overrepresentation analysis of genes positively (red) and negatively (blue) correlated with Stip1 . F Pearson’s correlation coefficient between Stip1 and key pluripotency factors: Pou5f1 ( R = 0.41), Sox2 ( R = 0.23) and Nanog ( R = 0.47).

Article Snippet: mESCs were derived from genetically modified mice expressing different levels of STIP1 or purchased from American Type Collection (ES-E14TG2a, ATCC #CRL-1821).

Techniques: Expressing

Journal: Communications Biology

Article Title: Stress-inducible phosphoprotein 1 (STIP1) is a critical stemness regulator in mouse embryonic stem cells and early mammalian development

doi: 10.1038/s42003-025-08763-9

Figure Lengend Snippet: A Schematic representation showing wild-type (WT), STIP1 +/− (50% expression) and STIP1 TgA (overexpression) mESCs lines derived from the inner cell mass of the blastocyst of genetically modified Stip1 mice. Total RNA extracted from these mESCs was subjected to bulk RNA sequencing followed by functional profiling. B Volcano plot of differentially expressed genes (DEGs) in STIP1 TgA versus (vs) WT. DEGs are classified as non-significant (gray), up- (red) or downregulated (blue), with genes of interest highlighted, p < 0.05. C Overrepresentation analysis (ORA) of Gene Ontology (GO) terms enriched among the DEGs of STIP1 TgA mESCs. D Volcano plot of non-significant (gray), up- (red) or downregulated (blue) DEGs of STIP1 +/− vs WT, p < 0.05. E ORA of GO terms enriched among the DEGs of STIP1 +/− , sorted by p -value. F Volcano plot of non-significant (gray), up- (red) and downregulated (blue) DEGs of STIP1 TgA vs STIP1 +/− , p < 0.05. G ORA of GO terms enriched among the DEGs of STIP1 TgA relative to STIP1 +/− , sorted by p -value.

Article Snippet: mESCs were derived from genetically modified mice expressing different levels of STIP1 or purchased from American Type Collection (ES-E14TG2a, ATCC #CRL-1821).

Techniques: Expressing, Over Expression, Derivative Assay, Genetically Modified, RNA Sequencing, Functional Assay

Journal: Communications Biology

Article Title: Stress-inducible phosphoprotein 1 (STIP1) is a critical stemness regulator in mouse embryonic stem cells and early mammalian development

doi: 10.1038/s42003-025-08763-9

Figure Lengend Snippet: A Gene expression (relative to WT) by RT-qPCR (2 -ΔΔCt method, Tbp and Gapdh as reference genes). One-way ANOVA, with Tukey’s post-hoc. #: p -values of STIP1 +/− or STIP1 TgA vs WT; *: p -values of STIP1 +/− vs STIP1 TgA . Stip1 ( # p = 0.0016, *** p = 0.0008), Pou5f1 / Oct4 (* p = 0.0187), Sox2 ( # p = 0.0150, ** p = 0.0011), Klf4 ( ### p = 0.0004, *** p = 0.0002), Nanog ( #### p < 0.0001 **** p < 0.0001), and Tfc2pl1 ( ### p = 0.0008, *** p = 0.0004). Mean ± standard deviation (SD), n = 3. B Representative western blot image for WT, STIP1 +/− , and STIP1 TgA mESCs genetically independent biological triplicates. C Protein expression (relative to WT) of STIP1 (* p = 0.0379, ** p = 0.0030, *** p = 0.0003), HSP70, OCT4 (* p = 0,0295, ** p = 0.0050), SOX2 (* p = 0.0173, ** p = 0.0036) and NANOG (* p = 0.0198, ** p = 0.0047), normalized by β-tubulin. One-way ANOVA, with Tukey’s post-hoc. Mean ± SD, n = 3. D Representative immunofluorescence panels of cells stained with STIP1 (green), SOX2 (yellow), and OCT4 (red). Merged images in the panel include DAPI-stained nuclei (blue). Scale bar = 10 µm. E Representative immunofluorescence panels of cells stained with DAPI/Nuclei (red) and SSEA1 (green). Scale bar = 10 µm. F Representative immunofluorescence panels of cells stained with DAPI/Nuclei (red) and Ki67 (green). Scale bar = 10 µm. G Percentage of Ki67 positive (Ki67+) mESCs. Welch’s ANOVA test with Dunnett’s T3 multiple comparisons test of WT ( n = 11), STIP1 +/− ( n = 12) and STIP1 TgA ( n = 14), *: p -values of STIP1 +/− or STIP1 TgA vs WT; #: STIP1 +/− vs STIP1 TgA , ** p = 0.0068, **** p < 0.0001, #### p < 0.0001. Mean ± SD. H Growth curve of WT, STIP1 +/− and STIP1 TgA mESCs. One-way ANOVA with Tukey’s post-hoc analysis was performed for each time-point. *: p -values of STIP1 +/− or STIP1 TgA vs WT; #: p -values of STIP1 +/− vs STIP1 TgA . 24 h : p = non-significant; 48 h : * p ≤ 0.0330, # p < 0.0001; 72 h : * p = 0.0340, ** p = 0.0062, # p < 0.0001; 96 h : * p = 0.0102, ** p = 0.0035, # p < 0.0001. Mean ± SD, n = 6. I Representative immunofluorescence panels of cells stained with DAPI/Nuclei (blue) and CC3 (red). Scale bar = 10 µm. J Quantification of the percentage of CC3 positive (CC3+) cells. Welch’s ANOVA test with Dunnett’s T3 multiple comparisons test of WT ( n = 29), STIP1 +/− ( n = 28) and STIP1 TgA ( n = 8). * p ≤ 0.0251, **** p < 0,0001, mean ± SD. K Representative immunofluorescence panels of cells stained with DAPI/Nuclei (blue) and pH2AX (green). Scale bar = 10 µm. L Quantification of the number of pH2AX foci each 10 µm. Welch’s ANOVA test with Dunnett’s T3 multiple comparisons test of WT ( n = 17), STIP1 +/− ( n = 12) and STIP1 TgA ( n = 11). * p = 0.0210; ** p = 0.0019, mean ± SD.

Article Snippet: mESCs were derived from genetically modified mice expressing different levels of STIP1 or purchased from American Type Collection (ES-E14TG2a, ATCC #CRL-1821).

Techniques: Gene Expression, Quantitative RT-PCR, Standard Deviation, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: Stress-inducible phosphoprotein 1 (STIP1) is a critical stemness regulator in mouse embryonic stem cells and early mammalian development

doi: 10.1038/s42003-025-08763-9

Figure Lengend Snippet: A Schematic diagram illustrating the generation of STIP1 ∆TPR1 mESCs lines derived from Stip1 genetically modified mice. B Volcano plot showing differentially expressed genes (DEGs) in STIP1 ∆TPR1 vs WT, with WT as the control group. Genes are classified as non-significant (gray), up- (red) or downregulated (blue), and DEGs of interest are highlighted. p < 0.05. C Overrepresentation analysis of Gene Ontology terms enriched among the DEGs of STIP1 ∆TPR1 mESCs. D Gene expression (relative to WT) by RT-qPCR (2 -ΔΔCt method, Gapdh as reference gene). Student’s t test for each individual gene. Stip1 - exons 7/9 (* p = 0.0189), Stip1 - exons 2/3 (*** p = 0.0002), Hsp90β , Pou5f1 / Oct4 , Sox2 (*** p = 0.0001), Nanog , Tfc2pl1 , Klf4 (** p = 0.0045), Stat3 and Esrrb (* p = 0.0316) and Ssea1 . Mean ± SD, n = 3. E Representative western blot image for WT and STIP1 ∆TPR1 mESCs for HSP90, STIP1, the truncated for of STIP1 (STIP1-∆TPR1) and Actin from genetically independent biological triplicates. F Protein expression (relative to WT) by band densitometry of HSP90 and STIP1, normalized by Actin. Student’s t test for each individual protein. ** p = 0.0012, * p = 0.0186. Mean ± SD, n = 3. G Representative western blot image for WT and STIP1 ∆TPR1 mESCs for OCT4, NANOG, SOX2 and β-tubulin from genetically independent biological triplicates. H Protein expression (relative to WT) by band densitometry of OCT4, NANOG, SOX2, normalized by β-tubulin. Student’s t test for each individual protein. *** p = 0.0006; * p ≤ 0.0204. Mean ± SD, n = 3. I Representative immunofluorescence panels of cells stained with STIP1 (green), OCT4 (red), and SOX2 (yellow). Merged images in the panel include DAPI-stained nuclei (blue). Scale bar = 10 µm. Scale bar = 10 µm. J Total immunofluorescence quantification (in arbitrary units) normalized by WT. Student’s t test for each individual protein relative to control was used for STIP1 and OCT4 expression. STIP1: *** p = 0.0002; OCT4: **p = 0.0020. Mann–Whitney test was used for SOX2 expression. SOX2: * p = 0.0411. Mean ± SD, n = 5. K Representative immunofluorescence panels of cells stained with DAPI/Nuclei (blue) and NANOG (red). Scale bar = 10 µm. Yellow arrowheads : cells in mitotic figures with observed NANOG reduction. L Representative immunofluorescence panels of cells stained with DAPI/Nuclei (red) and SSEA1 (green). Scale bar = 10 µm.

Article Snippet: mESCs were derived from genetically modified mice expressing different levels of STIP1 or purchased from American Type Collection (ES-E14TG2a, ATCC #CRL-1821).

Techniques: Derivative Assay, Genetically Modified, Control, Gene Expression, Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Staining, MANN-WHITNEY

Journal: Communications Biology

Article Title: Stress-inducible phosphoprotein 1 (STIP1) is a critical stemness regulator in mouse embryonic stem cells and early mammalian development

doi: 10.1038/s42003-025-08763-9

Figure Lengend Snippet: A Representative immunofluorescence panel of cells stained with DAPI/Nuclei (red) and Ki67 (green). Scale bar = 10 µm. B Quantification of Ki67-positive (Ki67+) mESCs. Student’s t test with Welch’s correction, **** p < 0.0001, WT: n = 85; STIP1 ΔTPR1 : n = 36. Mean ± SD. C Growth curve of WT and STIP1 ∆TPR1 mESCs was analyzed per day by Student’s t test. Day 1 (24 h after seeding): ** p = 0.0094; Day 3 (72 h): *** p = 0.0005 ; Day 5 (120 h): *** p = 0.0004; and Day 7 (168 h): *** p = 0.0004. Mean ± SD, n = 3. D Colorimetric analysis (corrected absorbance) of cell viability. Student’s t test, *** p = 0.009, Mean ± SD, n = 4. E Representative immunofluorescence panel of cells stained with DAPI/Nuclei (blue) and pH2AX (green). Scale bar = 10 µm. F Quantification of the number of pH2AX foci in each 10 µm. Student t test, *** p = 0.0009. Mean ± SD, n = 15. G Flow cytometry for WT and STIP1 ∆TPR1 shown as representative scatter plots, staining for BrdU (S-phase) and 7-AAD (total DNA content). H Quantification of the percentage of cells for each cell cycle phase. Student’s t test, ** p = 0.0085. Mean ± SD, n = 3. I Flow cytometry of cells labeled for RB1 ( p < 0.0001, all comparisons), p53 ( p < 0.0001, all comparisons), p21 ( p < 0.0001, WT vs STIP1 TgA , STIP1 ΔTPR1 vs STIP1 TgA ), CDC2 ( p < 0.0001, all comparisons), cyclin B1 ( p < 0.0001, all comparisons), and cyclin D1 ( p < 0.0001, all comparisons). One-way ANOVA with Tukey’s post-hoc. Between 13.000–30.000 events were measured, and the mean and standard deviation of those events were used for statistics. The color pattern represents the respective groups across all charts.

Article Snippet: mESCs were derived from genetically modified mice expressing different levels of STIP1 or purchased from American Type Collection (ES-E14TG2a, ATCC #CRL-1821).

Techniques: Immunofluorescence, Staining, Flow Cytometry, Labeling, Standard Deviation

Journal: Communications Biology

Article Title: Stress-inducible phosphoprotein 1 (STIP1) is a critical stemness regulator in mouse embryonic stem cells and early mammalian development

doi: 10.1038/s42003-025-08763-9

Figure Lengend Snippet: STIP1 forms a complex with HSP70 and HSP90 in the cytoplasm of different cell types, assisting in proteostasis. STIP1 knockdown (STIP1 +/− ) in mESCs results in decreased (blue arrowheads) proliferation and increased (red arrowheads) apoptosis. Furthermore, nuclear localization of the protein can be observed, concomitant with reduced expression of the pluripotency marker SOX2, and increased DNA damage. Low expression levels of a truncated form of the protein (STIP1 ∆TPR1 ) recapitulate some of these phenotypes, including increased DNA damage, decreased SOX2, decreased cell proliferation and viability. We also observed altered cell cycle profile in STIP1 ∆TPR1 mESCs, which presented G2 arrest. Increased expression of STIP1 (STIP1 TgA ) leads to increased proliferation, decreased apoptosis and DNA damage and increased expression of pluripotency markers. In cells prone to differentiation, we see a strong expression of STIP1 in actin filaments, and a preferential localization of STIP1 in the nucleus in all studied cell lines, but more frequently in STIP1 +/− and STIP1 ∆TPR1 . This could indicate a location- and stage-specific function of STIP1 in the process of differentiation. In accordance, STIP1 decrease in STIP1 +/− and STIP1 ∆TPR1 results in impaired embryoid body formation, while we propose that increased STIP1 leads to late differentiation as it increases the expression of pluripotency markers in mESCs.

Article Snippet: mESCs were derived from genetically modified mice expressing different levels of STIP1 or purchased from American Type Collection (ES-E14TG2a, ATCC #CRL-1821).

Techniques: Knockdown, Expressing, Marker

Journal: Exploration

Article Title: STIP1 drives Metabolic Reprogramming in Esophageal Squamous Cell Carcinoma via AHCY‐LDHA Axis

doi: 10.1002/exp.20240198

Figure Lengend Snippet: FIGURE 1 Heat-stimulation promotes glycolysis and induces STIP1 overexpression. A. WB experiment was performed to measure the glycolysis enzymes level. B. Glucose uptake was detected by kit after 65◦C water stimulation. C. STIP1 expression was heightened in esophageal tissues of mice treated with hot water compared to normal controls. D. Across a panel of human esophageal cell lines, STIP1 protein levels were elevated in multiple ESCC lines versus normal esophageal epithelial cells (upper panel). The bands intensities were subjected to quantitative analysis (lower panel). E. In analysis of 5 paired patient ESCC tumor and adjacent normal tissues, STIP1 protein was increased in tumor samples compared to matched normal epithelium (upper panel). The bands intensities were subjected to quantitative analysis (lower panel). TCGA database was used to evaluate STIP1 expression across normal and cancerous tissues (F), examining its distribution across tumor stages (G) and investigating correlations between STIP1 expression and patient prognostic outcomes (H). I. By immunohistochemistry of ESCC tissue microarrays, STIP1 staining was more intense in tumor areas versus paired normal epithelium (40x and 100x magnifications shown). Quantitative analysis confirmed significantly higher STIP1 protein levels in ESCC tumors compared to paired normal tissues(J) and in unpaired ESCC versus normal esophagus (K). Stratifying by clinicopathologic characteristics, STIP1 expression was incrementally increased with higher lymph node involvement (L), poorer tumor grade (M), and advanced tumor stage (N). O. Relationship between STIP1 expression level and overall survival for tissue microarray. P. Schematic of the 4NQO-induced esophageal cancer model.

Article Snippet: Cells were then incubated overnight at 4◦C with primary antibodies targeting STIP1 (Proteintech, 68155-1-Ig, 1:200 dilution) and AHCY (Proteintech, 10757-2-AP, 1:100 dilution).

Techniques: Over Expression, Expressing, Immunohistochemistry, Staining, Microarray

Journal: Exploration

Article Title: STIP1 drives Metabolic Reprogramming in Esophageal Squamous Cell Carcinoma via AHCY‐LDHA Axis

doi: 10.1002/exp.20240198

Figure Lengend Snippet: FIGURE 2 STIP1 promotes cell growth and PDXO development. A. Stable STIP1 knockdown ESCC cell lines were generated, with efficient STIP1 protein reduction confirmed by western blotting. B. STIP1 knockdown significantly decreased ESCC cell proliferation based on MTT assays. C. D Anchorage independent growth and plate colony formation assay from different cells with STIP1 knockdown. Colonies were counted using Image Pro- Plus (Scale bar: 200 µm). E. STIP1 was overexpressed in KYSE410 cells and the expression of STIP1 was determined by western blot. F. After overexpressed STIP1, cell proliferation was measured by MTT assay. G, H. Anchorage independent growth and plate colony formation assay from KYSE410 with STIP1 overexpression. Colonies were counted using Image J-Plus (Scale bar: 200 µm). I-J Esophageal squamous cell carcinoma cells with stable knockdown of STIP1 expression via short hairpin RNAs (shRNAs) or non-targeting control shRNA were subcutaneously injected into athymic nude mice (n = 8 per group) to evaluate effects on tumorigenicity. Tumor weights were calculated in the right panel. K-L. Intertumoral injection of STIP1 shRNA in PDXs suppressed tumor growth compared to control shRNA. Tumor weights were calculated in the right panel. M-N. STIP1 knockdown reduced the number and size of PDXOs. Organoids absorbance was measured in the right panel. All data statistical differences were evaluated using Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Cells were then incubated overnight at 4◦C with primary antibodies targeting STIP1 (Proteintech, 68155-1-Ig, 1:200 dilution) and AHCY (Proteintech, 10757-2-AP, 1:100 dilution).

Techniques: Knockdown, Generated, Western Blot, Colony Assay, Expressing, MTT Assay, Over Expression, Control, shRNA, Injection

Journal: Exploration

Article Title: STIP1 drives Metabolic Reprogramming in Esophageal Squamous Cell Carcinoma via AHCY‐LDHA Axis

doi: 10.1002/exp.20240198

Figure Lengend Snippet: FIGURE 3 STIP1 plays a crucial role in glycolysis. A. Western blot shows STIP1 knockdown in KYSE450 and KYSE30 cells reduce protein levels of key glycolytic enzymes including PKM2, PKM, LDHA, ENO1 and ALDOA in ESCC cells. β-actin is shown as a loading control. B. STIP1 knockdown decreases glucose consumption in ESCC cells. Values are normalized to total protein content. C. STIP1 knockdown reduces lactate secretion in ESCC cells. Lactate levels in conditioned media were normalized to cell number. D. STIP1 knockdown lowers LDHA catalytic activity in ESCC cell lysates. E. Overexpression STIP1 enhance LDHA enzyme activity. LDHA activity was determined by measuring NADH levels spectrophotometrically. F,G. ECAR reflecting glycolytic flux, is decreased or increased after STIP1 knockdown in KYSE450 or overexpression in KYSE410 cells. ECAR was measured using a Seahorse XFe96 analyzer under basal conditions and in response to glucose, oligomycin, and 2-DG. Data are mean ± SD of triplicate experiments. *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t-test.

Article Snippet: Cells were then incubated overnight at 4◦C with primary antibodies targeting STIP1 (Proteintech, 68155-1-Ig, 1:200 dilution) and AHCY (Proteintech, 10757-2-AP, 1:100 dilution).

Techniques: Western Blot, Knockdown, Control, Activity Assay, Over Expression

Journal: Exploration

Article Title: STIP1 drives Metabolic Reprogramming in Esophageal Squamous Cell Carcinoma via AHCY‐LDHA Axis

doi: 10.1002/exp.20240198

Figure Lengend Snippet: FIGURE 4 STIP1 interacts with AHCY to enhance binding to LDHA, thereby promoting glycolytic flux. A. Mass spectrometry analysis identifies glycolytic enzymes as STIP1-interacting proteins. B, C. Endogenous and exogenous co-immunoprecipitation verifies binding between STIP1 and AHCY D. Immunofluorescence staining shows co-localization of STIP1 (green) and AHCY (red) in the cytoplasm of ESCC cells (DAPI stain in blue). E. The AHCY-STIP1 complex was modeled using Alphafold3. Then the AHCY in the model was aligned to the experimental structure (PDB 1LI4), and colored by C-alpha RMSD using ChimeraX. Red indicates high RMSD and larger conformational change. F. Knockdown or overexpression of STIP1 reduces or enhances AHCY enzyme activity, respectively. G,H. Co-immunoprecipitation confirms endogenous and exogenous interaction between AHCY and LDHA. I. Endogenous IP assay to check the binding of STIP1 and LDHA. J. HEK293T cells were transfected with Myc-tagged AHCY and truncated LDHA including 1–274, 1–160, 161–274. Immunoprecipitation was performed using anti-HA affinity gel followed by western blots using anti-Myc antibody. K. Immunofluorescence shows cytoplasmic co-localization of AHCY (green) and LDHA (red) in ESCC cells. L,M. STIP1 knockdown or overexpression reduces or enhances the AHCY-LDHA interaction. N,O. Knockdown or overexpression of STIP1 or AHCY decreases or increases the SAM:SAH ratio. Data are mean ± SD of 3 experiments. *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t-test.

Article Snippet: Cells were then incubated overnight at 4◦C with primary antibodies targeting STIP1 (Proteintech, 68155-1-Ig, 1:200 dilution) and AHCY (Proteintech, 10757-2-AP, 1:100 dilution).

Techniques: Binding Assay, Mass Spectrometry, Immunoprecipitation, Immunofluorescence, Staining, Knockdown, Over Expression, Activity Assay, Transfection, Western Blot

Journal: Exploration

Article Title: STIP1 drives Metabolic Reprogramming in Esophageal Squamous Cell Carcinoma via AHCY‐LDHA Axis

doi: 10.1002/exp.20240198

Figure Lengend Snippet: FIGURE 7 LCA is a STIP1 inhibitor. A. Chemical structure of LCA. B. Computational modeling predicts binding between LCA and STIP1. C. Pull- down assay shows LCA directly interacts with STIP1 protein in vivo. D. SPR analysis verifies direct binding between LCA and STIP1. Sensorgram shows response over time as LCA passes over STIP1-conjugated chip. E. Cellular thermal shift assay demonstrates LCA binds and stabilizes STIP1 protein in ESCC cells. F. Overview of quantitative proteomics approach. KYSE450 cells were treated with LCA or DMSO control for 24 h, followed by tandem mass tag (TMT) labeling and LC-MS/MS analysis. G. After treat various doses of LCA, AHCY, LDHA, PKM2 and ALDOA protein level were measured by western blot. H,I. LCA decreases glucose consumption and LDH vitality in ESCC cells. J,K. ECAR is lowered by LCA, reflecting impaired glycolysis. Data are mean ± SD of 3 experiments. *p < 0.05, **p < 0.01, by Student’s t-test.

Article Snippet: Cells were then incubated overnight at 4◦C with primary antibodies targeting STIP1 (Proteintech, 68155-1-Ig, 1:200 dilution) and AHCY (Proteintech, 10757-2-AP, 1:100 dilution).

Techniques: Binding Assay, Pull Down Assay, In Vivo, Thermal Shift Assay, Quantitative Proteomics, Control, Labeling, Liquid Chromatography with Mass Spectroscopy, Western Blot

Journal: International journal of biological sciences

Article Title: Macrophage-derived S100A9 promotes diabetic cardiomyopathy by disturbing mitochondrial quality control via STAT3 activation.

doi: 10.7150/ijbs.111128

Figure Lengend Snippet: Figure 7. S100A9 promotes STIP1-STAT3 interaction in cardiomyocytes and diabetic heart tissues. a, Upset-Venn plot of IP-MS results in rhS100A9-treated AC16 cardiomyocytes. b-c, co-immunoprecipitation (co-IP) of MYC-STAT3 and HA-STIP1 in AC16 cardiomyocytes. d, S100A9 promoted STIP1 binding with STAT3 in AC16 cells transfected with Flag-S100A9, MYC-STAT3 and HA-STIP1. e, The predicted binding sites between STIP1 and STAT3. Optimized pose model of STIP1-STAT3 interaction by R-Dock analysis. f, Full-length STIP1 or truncated mutant STIP1 (TPR5-8 domain, Δ259-427) were co-transfected with MYC-STAT3 in AC16 cardiomyocytes. co-IP of STAT3 and STIP1 was perform with HA-tag antibody. g, co-IP of STAT3 and STIP1 was performed in STZ-induced or db/db diabetic heart tissues treated with/without paquinimod (PAQ).

Article Snippet: S100A8 (15792-1-AP), S100A9 (26992-1-AP), STIP1 (15218-1-AP), FIS1 (10956-1-AP), Tubulin (10068-1-AP), Lamin B (12987-1-AP), GAPDH (10494-1-AP), and HA-tag (51064-2-AP) were obtained from Proteintech Technology (Proteintech, China).

Techniques: Protein-Protein interactions, Immunoprecipitation, Co-Immunoprecipitation Assay, Binding Assay, Transfection, Mutagenesis