Journal: Nature Communications

Article Title: Cell-type-specific functionality encoded within the intrinsically disordered regions of OCT4

doi: 10.1038/s41467-025-63806-3

Figure Lengend Snippet: a Proteins associated with OCT4 WT or mutants on chromatin identified by ChIP-SICAP rank ordered by normalized signal intensity from two biological replicates ( n = 2). Top ranked proteins and TNIP2 are highlighted. b Volcano plot illustrating the differentially enriched proteins co-localised with OCT4 in early reprogramming versus ESCs. Statistical significance assessed by unpaired t -test and FDR-adjusted P value. The −log10 ( P value) is plotted against the log2 fold change. The significance threshold is represented by the non-axial lines denoting; |log2(fold change)| > 1.5 and FDR = 0.1. OCT4 partners enriched uniquely in ESCs (green squares), shared but more enriched in ESCs (blue squares), enriched uniquely in early reprogramming (red circles), shared but more enriched in early reprogramming (purple circles), or shared and equally enriched (grey squares). Example proteins are indicated. c Heatmaps showing the differential abundance of OCT4-interacting proteins as measured by SILAC-MS and expressed as a log2 hFibs/hESCs ratio (colour intensity scale at the bottom) from n = 3 biological replicates. Examples of OCT4 partners and number of proteins ( n ) are indicated. d Heatmap showing hierarchal clustering of proteins identified by ChIP-SICAP for OCT4 WT and mutant in hESCs and early reprogramming. Examples from selected groups are highlighted. Number of proteins ( n ) identified in each group is indicated. The colour intensity scale is shown at the bottom. Source data are provided as a Source Data file.

Article Snippet: The primary antibody incubations with anti-human OCT4 antibody (0.45 mg/ml #ab19857; Abcam), anti-human GAPDH (1:5000 dilution of GTX627408; GeneTex Inc), anti-human TNIP2 (1:1000; #15459-1-AP, ProteinTech), anti-human ETV4 (1:1000; #PA5-79223, ThermoScientific), anti-human XPO6 (1 μg/ml, #ab72333, Abcam), anti-human UFD1L (1:1000; #10615-1-AP, ProteinTech), anti-human histone H3 (1:10,000; #ab24834, Abcam) and anti-human lamin (0.5 μg/mL; #ab16048, Abcam) were performed at 4 °C for 16 h. The secondary antibody incubations with goat anti-rabbit IgG-HRP (1:5000; #sc-2004, Santa Cruz Biotechnology) and goat anti-mouse IgG-HRP (1:5000; #sc-2005 Santa Cruz Biotechnology) were performed for 1 h at room temperature.

Techniques: Multiplex sample analysis, Mutagenesis

Journal: Nature Communications

Article Title: Cell-type-specific functionality encoded within the intrinsically disordered regions of OCT4

doi: 10.1038/s41467-025-63806-3

Figure Lengend Snippet: a Schematic of reprogrammable MEFs (cas9-TNG-MKOS-MEFs), expressing GFP under Nanog promoter, cas9 under Elf-a promoter, and dox-inducible MKOS-ires-mOrange. b Experimental scheme for reprogramming cas9-TNG-MKOS-MEFs to iPSCs after targeting specific genes with sgRNA. c Whole-well images showing GFP (green) and RFP fluorescence (red) colonies after gRNA targeting (indicated on top). d Bar plots of GFP positive iPSC colonies shown in ( c ). e Western blots of OCT4 partners in hESCs, OSKM-48h, and hFibs. Molecular weight standards (kDa) are indicated. Histone H3 was used as loading control. f Immunofluorescence showing the nuclear localisation (green) of TNIP2CT and TNIP2CT-OCT4 fusion compared to TNIP2 (protein schematics shown above). DAPI (blue) and CellMask (red) staining were used to indicate the nucleus and the plasma membrane, respectively. Scalebar, 100 µm. g Whole-well images showing DAPI (blue), NANOG (red) and merged (magenta) of iPSCs generated by TNIP2CT-OCT4 fusion derivatives compared to OCT4 WT and lin29-42 mutant. Scalebar, 5 mm. h Bar plots of NANOG +ve colonies (purple) versus DAPI (blue) generated by OCT4 derivative shown in ( g ). i Experimental flowchart of ATAC-seq during the early reprogramming after knocking down TNIP2 using siRNA. STAT3 and non-targeting siRNA were used as controls. j Western blots confirming diminished protein levels of TNIP2 (top) and STAT3 (bottom) after siRNA-KD in day 3 and 6 as shown in ( i ). k ATAC-seq read density heatmaps spanning ±1 kb from the centre of OCT4 peaks identified in (Fig. ) during early reprogramming and after TNIP2-KD and STAT3-KD as shown in ( i ). Average profile plots are shown to the left. Number ( n ) of OCT4 sites and ATAC enrichment scale (RPGC) are indicated. Images are representatives of n = 3 ( c , f , and j ) and n = 6 ( g ) biological replicates. Data are presented as mean values +/- SEM of n = 3 ( d ) and n = 6 ( h ) biological replicates. Statistical significance assessed by one-way ANOVA ( d ) or unpaired t -test ( h ) as indicated by adjusted P values (n.s > 0.05, *≤ 0.05, **≤ 0.01, ***≤ 0.001, ****≤ 0.0001). Source data are provided as a Source Data file.

Article Snippet: The primary antibody incubations with anti-human OCT4 antibody (0.45 mg/ml #ab19857; Abcam), anti-human GAPDH (1:5000 dilution of GTX627408; GeneTex Inc), anti-human TNIP2 (1:1000; #15459-1-AP, ProteinTech), anti-human ETV4 (1:1000; #PA5-79223, ThermoScientific), anti-human XPO6 (1 μg/ml, #ab72333, Abcam), anti-human UFD1L (1:1000; #10615-1-AP, ProteinTech), anti-human histone H3 (1:10,000; #ab24834, Abcam) and anti-human lamin (0.5 μg/mL; #ab16048, Abcam) were performed at 4 °C for 16 h. The secondary antibody incubations with goat anti-rabbit IgG-HRP (1:5000; #sc-2004, Santa Cruz Biotechnology) and goat anti-mouse IgG-HRP (1:5000; #sc-2005 Santa Cruz Biotechnology) were performed for 1 h at room temperature.

Techniques: Expressing, Fluorescence, Western Blot, Molecular Weight, Control, Immunofluorescence, Staining, Clinical Proteomics, Membrane, Generated, Mutagenesis

Journal: Immunity, inflammation and disease

Article Title: TNFAIP3 interacting protein 2 relieves lipopolysaccharide (LPS)-induced inflammatory injury in endometritis by inhibiting NF-kappaB activation.

doi: 10.1002/iid3.970

Figure Lengend Snippet: FIGURE 1 The TNIP2 expression level in LPS‐induced hEECs. (A) Western blot assay was performed to quantify the patterns of TNIP2. (B) The TNIP2 mRNA level in LPS‐induced hEECs. Data are exhibited as average ± SD of triple single experiments. N = 3. **p < .01 versus Control. hEECs, human endometrial epithelial cells; LPS, lipopolysaccharide; SD, standard deviation; TNIP2, TNFAIP3 interacting protein 2.

Article Snippet: The PVDF membrane was then incubated for 12 h with primary antibodies (Proteintech) against TNIP2 (15459‐1‐AP; Proteintech), Caspase3 (19677‐1‐AP; Proteintech), p65 NF‐κB (66535‐1‐Ig; Proteintech), and phospho‐p65 NF‐ κB (ARG51518; Arigo).

Techniques: Expressing, Western Blot, Control, Standard Deviation

Journal: Immunity, inflammation and disease

Article Title: TNFAIP3 interacting protein 2 relieves lipopolysaccharide (LPS)-induced inflammatory injury in endometritis by inhibiting NF-kappaB activation.

doi: 10.1002/iid3.970

Figure Lengend Snippet: FIGURE 2 Effect of TNIP2 on apoptosis of LPS‐induced hEECs. (A) The efficiency of TNIP2‐plasmid transfection was determined by western blot assay. (B) The efficiency of TNIP2‐plasmid transfection was determined by RT‐qPCR. (C) CCK‐8 was carried out to calculate cell viability. (D) The LDH detection kit was used to measure LDH activity. (E and F) The FCM assay was carried out to assess cell apoptosis. (G) The capase3 activity detection kit was carried out to measure caspase3 activity. (H and I) Western blot analysis was performed to quantify the patterns of apoptosis‐related protein (cleaved‐caspase3). Data are exhibited as average ± SD of triple single experiments. N = 3. **p < .01 versus Control‐plasmid; ##p < .01 versus Control; &&p < .01 versus LPS + control‐plasmid. CCK‐8, cell counting kit‐8; FCM, flow cytometry; hEECs, human endometrial epithelial cells; LDH, lactate dehydrogenase; LPS, lipopolysaccharide; SD, standard deviation; TNIP2, TNFAIP3 interacting protein 2.

Article Snippet: The PVDF membrane was then incubated for 12 h with primary antibodies (Proteintech) against TNIP2 (15459‐1‐AP; Proteintech), Caspase3 (19677‐1‐AP; Proteintech), p65 NF‐κB (66535‐1‐Ig; Proteintech), and phospho‐p65 NF‐ κB (ARG51518; Arigo).

Techniques: Plasmid Preparation, Transfection, Western Blot, Quantitative RT-PCR, CCK-8 Assay, Activity Assay, Control, Cell Counting, Flow Cytometry, Standard Deviation

Journal: Immunity, inflammation and disease

Article Title: TNFAIP3 interacting protein 2 relieves lipopolysaccharide (LPS)-induced inflammatory injury in endometritis by inhibiting NF-kappaB activation.

doi: 10.1002/iid3.970

Figure Lengend Snippet: FIGURE 3 Impact of TNIP2 on inflammation and oxidative stress of LPS‐induced hEECs. (A–C) The concentration of TNF‐α, IL‐1β, and IL‐6 was detected by ELIZA. (D) ROS assay kit was performed to confirm the ROS release. (E and F) The SOD and CAT activity kits were enacted to detect the activity of SOD and CAT. Data are exhibited as average ± SD of triple single experiments. N = 3. **p < .01 versus Control; ##p < .01 versus LPS + control‐plasmid. CAT, catalase; ELIZA, enzyme‐linked‐immunosorbent analysis; hEECs, human endometrial epithelial cells; IL, Interleukin; LPS, lipopolysaccharide; ROS, reactive oxygen species; SD, standard deviation; SOD, superoxide dismutase; TNF‐α, tumor necrosis factor‐alpha; TNIP2, TNFAIP3 interacting protein 2.

Article Snippet: The PVDF membrane was then incubated for 12 h with primary antibodies (Proteintech) against TNIP2 (15459‐1‐AP; Proteintech), Caspase3 (19677‐1‐AP; Proteintech), p65 NF‐κB (66535‐1‐Ig; Proteintech), and phospho‐p65 NF‐ κB (ARG51518; Arigo).

Techniques: Concentration Assay, ROS Assay, Activity Assay, Control, Plasmid Preparation, Standard Deviation

Journal: Immunity, inflammation and disease

Article Title: TNFAIP3 interacting protein 2 relieves lipopolysaccharide (LPS)-induced inflammatory injury in endometritis by inhibiting NF-kappaB activation.

doi: 10.1002/iid3.970

Figure Lengend Snippet: FIGURE 6 NF‐κB agonist CU‐T12‐9 reversed TNIP2‐induced apoptosis reduction in LPS‐induced hEECs. (A) CCK‐8 was presented to obtain cell viability. (B) The LDH detection kit was carried out to measure LDH activity. (C and D) The apoptosis rate in hEECs was confirmed by FCMs assay. (E) The capase3 activity detection kit was used to measure caspase3 activity in hEECs. (F) The cleaved‐caspase3 protein expression was verified by western blot assay. (G) Cleaved‐caspase3/GAPDH ratio. Data are exhibited as average ± SD of triple single experiments. N = 3. **p < .01 versus LPS + control‐plasmid; ##p < .01 versus LPS + TNIP2‐plasmid. CCK‐8, cell counting kit‐8; FCM, flow cytometry; hEECs, human endometrial epithelial cells; LDH, lactate dehydrogenase; LPS, lipopolysaccharide; NF‐κB, nuclear factor‐kappa; SD, standard deviation; TNIP2, TNFAIP3 interacting protein 2.

Article Snippet: The PVDF membrane was then incubated for 12 h with primary antibodies (Proteintech) against TNIP2 (15459‐1‐AP; Proteintech), Caspase3 (19677‐1‐AP; Proteintech), p65 NF‐κB (66535‐1‐Ig; Proteintech), and phospho‐p65 NF‐ κB (ARG51518; Arigo).

Techniques: CCK-8 Assay, Activity Assay, Expressing, Western Blot, Control, Plasmid Preparation, Cell Counting, Flow Cytometry, Standard Deviation

Journal: Immunity, inflammation and disease

Article Title: TNFAIP3 interacting protein 2 relieves lipopolysaccharide (LPS)-induced inflammatory injury in endometritis by inhibiting NF-kappaB activation.

doi: 10.1002/iid3.970

Figure Lengend Snippet: FIGURE 5 NF‐κB agonist CU‐T12‐9 activated NF‐kB pathway in LPS‐induced hEECs. (A) The p‐p65 and p65 levels were estimated by western blot analysis. (B) The ratio of p‐p65/p65. Data are exhibited as average ± SD of triple single experiments. N = 3. **p < .01 versus LPS + control‐plasmid; ##p < .01 versus LPS + TNIP2‐plasmid. hEECs, human endometrial epithelial cells; LPS, lipopolysaccharide; NF‐κB, nuclear factor‐kappa; SD, standard deviation; TNIP2, TNFAIP3 interacting protein 2.

Article Snippet: The PVDF membrane was then incubated for 12 h with primary antibodies (Proteintech) against TNIP2 (15459‐1‐AP; Proteintech), Caspase3 (19677‐1‐AP; Proteintech), p65 NF‐κB (66535‐1‐Ig; Proteintech), and phospho‐p65 NF‐ κB (ARG51518; Arigo).

Techniques: Western Blot, Control, Plasmid Preparation, Standard Deviation

Journal: Immunity, inflammation and disease

Article Title: TNFAIP3 interacting protein 2 relieves lipopolysaccharide (LPS)-induced inflammatory injury in endometritis by inhibiting NF-kappaB activation.

doi: 10.1002/iid3.970

Figure Lengend Snippet: FIGURE 7 NF‐κB agonist CU‐T12‐9 reversed the TNIP2‐induced inflammation and oxidative stress suppression in LPS‐induced hEECs. (A–C) The concentration of TNF‐α, IL‐1β, and IL‐6 was detected by ELIZA. (D) The ROS kit was used to confirm the ROS release. (E and F) SOD and CAT activity kits were enacted to detect the activity of SOD and CAT. Data are exhibited as average ± SD of triple single experiments. N = 3. **p < .01 versus LPS + control‐plasmid; ##p < .01 versus LPS + TNIP2‐plasmid. CAT, catalase; ELIZA, enzyme‐linked‐ immunosorbent analysis; hEECs, human endometrial epithelial cells; IL, Interleukin; LPS, lipopolysaccharide; NF‐κB, nuclear factor‐kappa; ROS, reactive oxygen species; SD, standard deviation; SOD, superoxide dismutase; TNF‐α, tumor necrosis factor‐alpha; TNIP2, TNFAIP3 interacting protein 2.

Article Snippet: The PVDF membrane was then incubated for 12 h with primary antibodies (Proteintech) against TNIP2 (15459‐1‐AP; Proteintech), Caspase3 (19677‐1‐AP; Proteintech), p65 NF‐κB (66535‐1‐Ig; Proteintech), and phospho‐p65 NF‐ κB (ARG51518; Arigo).

Techniques: Concentration Assay, Activity Assay, Control, Plasmid Preparation, Standard Deviation