Journal: Nature Communications

Article Title: Metal-organic polyhedra maintain the self-renewal of embryonic stem cells

doi: 10.1038/s41467-025-63811-6

Figure Lengend Snippet: A The photo of MOP-1 aqueous solution (left). Monodisperse structure of MOP-1 in H 2 O (right). Color codes: V, green; O, red; C, gray; N, blue; Cl, bright blue. The large pink sphere represents the free space inside the molecular cage. For clarity, H atoms were omitted. Schematic (right panel) was created with Diamond software. B Experimental and simulated PXRD patterns of MOP-1. C TEM image of MOP-1 (left). Scale bar, 20 nm. The particle size of MOP-1 in H 2 O (right). Three experiments were repeated independently with similar results. D The schematic diagram showed that the SHP-2 mediated STAT3 inactivation. Schematic diagram was created with Microsoft Office PowerPoint. E , F An in-depth mechanism investigation of mESC pluripotency control by MOP-1. Binding model from a global view of a complex composed of SHP-2 and MOP-1 illustrated by electrostatic surface potential ( E ). Binding modes are illustrated by ribbon diagrams of a complex composed of SHP-2 and MOP-1 (the left panel), a complex composed of SHP-2 with MOP (the middle panel) and a complex composed of SHP-2 with ZrMOP (the right panel). The top panel is the global view of the catalytic PTP structure of SHP-2, the bottom panel is the focused view of binding modes illustrated by the Ribbon diagrams ( F ). G Binding kinetics of MOP-1 (top panel) and MOP-2 (bottom panel) with SHP-2 were measured by the SPR assay. H The ICP-MS analysis of the binding quantity between MOPs and SHP-2 (mean ± s.e.m, n = 6). I The inhibition efficiency of SHP-2, JAK2, JAK1, SHP-1, PTP1B, Cyt c , ACP and lipase by MOP-1 at a concentration of 2 μM using enzyme assay (mean ± s.e.m, n = 3). Data in ( H ) and ( I ) are analyzed by one-way ANOVA. **** P < 0.0001, the binding between MOP-1 and SHP-2 vs. the binding between MOP-2 and SHP-2, relative activity of SHP-2 vs. relative activity of JAK2, JAK1, PTP1B, Cyt c , ACP and lipase. *** P < 0.001, relative activity of SHP-2 vs. relative activity of SHP-1.

Article Snippet: The binding kinetics and affinity of MOP-1 or MOP-2 to SHP-2 (MCE, HY-P700618), SHP-1 (MCE, HY- P71141 ), JAK1 (MCE, HY-P700583), JAK2 (MCE, HY-P701102) and PTP1B (MCE, HY- P73685 ) were analyzed by SPR (Biacore 8 K, Cytiva).

Techniques: Software, Control, Binding Assay, SPR Assay, Inhibition, Concentration Assay, Enzymatic Assay, Activity Assay

Journal: bioRxiv

Article Title: Spatial mapping of proteins and their activity states in cancer models by multiplex in situ PLA

doi: 10.1101/2025.07.11.662357

Figure Lengend Snippet: misPLA analyses of phosphorylation states and protein-protein interactions in SK-BR cells with or without EGF stimulation. A) Phosphorylation targets: SK-BR cells were analyzed for phosphorylation of STAT5a (pSTAT5a), STAT3 (pSTAT3), AKT (pAKT), ERK (pERK), and EGFR (pEGFR), under unstimulated and EGF-stimulated conditions. All targets were visualized three at a time in sequential detection cycles and are shown simultaneously (upper panels, “All targets”) and subsequently as individual channels. PLA signals (red, cyan, green, purple) reflect activated protein states detected via dual-recognition proximity ligation. DAPI (blue) labels nuclei. Scale bars = 50 µm. B) Protein-protein interactions: Visualization of the following protein-protein interactions investigated by misPLA in unstimulated and EGF-stimulated SK-BR cells: MEK1–ERK2, GRB2–MEK1, EGFR–GRB2, STAT3–STAT5a, JAK1–JAK3, JAK1–PI3K, JAK2–STAT5a, JAK1–STAT3, and JAK2–JAK3. Upper panels (“All targets”) represent simultaneous visualization of all targets, imaged three at a time in sequential detection cycles, followed by separated signals per interaction. Scale bars = 50 µm.

Article Snippet: The following primary antibodies were used for Western blotting: JAK1 (ProteinTech, 66466-1-Ig), STAT3 (ProteinTech, 60199-1-Ig; Abcam, ab171359), MEK1 (Abcam, ab239802), EGFR (Abcam, ab271834), AKT2 (Thermo Scientific, PA5-85518), ERK2 (Thermo Fisher, PA5-29636), phospho-PI3K p85/p55 (Cell Signaling Technology, 4228S), pSTAT3-Y705 (R&D Systems, AF4607), Grb2 (R&D Systems, mab38461), GAPDH (CST, 14C10), and Vinculin (CST, E1E9V), used at either 1:1000 or 1:2000 dilution.

Techniques: Phospho-proteomics, Protein-Protein interactions, Ligation

Journal: bioRxiv

Article Title: Spatial mapping of proteins and their activity states in cancer models by multiplex in situ PLA

doi: 10.1101/2025.07.11.662357

Figure Lengend Snippet: misPLA mapping of signaling interactions in a lymph-node Hodgkin lymphoma, mixed cellularity (right neck); Hodgkin lymphoma, lymphocyte-depleted (neck); Hodgkin lymphoma, lymphocyte-predominant (left neck); Hodgkin lymphoma, mixed cellularity (left neck); and thymoma type B3 (mediastinum). Top row (visualization cycle 1) displays MEK1–ERK2 (FITC), EGFR–GRB2 (Cy5) and GRB2–MEK1 (Cy3N) together with DAPI. Middle row (cycle 2) shows STAT3–STAT5a (FITC), JAK1–JAK3 (Cy5) and JAK1–PI3Kp85 (Cy3N). Bottom row (cycle 3) presents JAK2– STAT5a (FITC), JAK2–JAK3 (Cy5) and JAK1–STAT3 (Cy3N). All nine pairs of antibody-oligonucleotide conjugates were applied and then amplified in a single incubation. The RCA products were revealed using detection oligonucleotides conjugated with three fluorophores in three visualization cycles. A standard three-channel fluorescence microscope was used with identical settings for all three fluorophores. Scale bars, 50 µm.

Article Snippet: The following primary antibodies were used for Western blotting: JAK1 (ProteinTech, 66466-1-Ig), STAT3 (ProteinTech, 60199-1-Ig; Abcam, ab171359), MEK1 (Abcam, ab239802), EGFR (Abcam, ab271834), AKT2 (Thermo Scientific, PA5-85518), ERK2 (Thermo Fisher, PA5-29636), phospho-PI3K p85/p55 (Cell Signaling Technology, 4228S), pSTAT3-Y705 (R&D Systems, AF4607), Grb2 (R&D Systems, mab38461), GAPDH (CST, 14C10), and Vinculin (CST, E1E9V), used at either 1:1000 or 1:2000 dilution.

Techniques: Amplification, Incubation, Fluorescence, Microscopy

Journal: Molecular Cancer Research

Article Title: IFI16 in Human Prostate Cancer

doi: 10.1158/1541-7786.mcr-06-0269

Figure Lengend Snippet: FIGURE 3. TSA-mediated increased expression of IFI16 in LNCaP cells is not dependent on increased expression of JAK1 protein kinase. A. Total cell extracts prepared from control (lane 1) LNCaP cells or cells treated with 5-aza-dC (5-AZA; 5 Amol/L; lane 2), TSA (100 nmol/L; lane 3), or both 5-aza-dC and TSA together (lane 4) for 24 h were analyzed by immunoblotting using antibodies specific to the indicated proteins. The treatment-induced fold changes in JAK1 protein levels are at the bottom of the figure. B. LNCaP cells were either nucleofected with an empty vector (lanes 1 and 3) or a plasmid encoding human JAK1 protein kinase (lanes 2 and 4) as described in Materials and Methods. Twenty- four hours after nucleofections, cells were either left untreated (lanes 1 and 2) or treated with IFNs (IFN-a, 1,000 units/mL; IFN-g, 20 ng/mL; lanes 3 and 4). Forty-eight hours after nucleofections, total cell lysates were analyzed by immunoblotting using antibodies specific to the indicated proteins. C. Total cell extracts prepared from control (lane 1) LNCaP cells or cells treated with IFNs (IFN-a, 1,000 units/mL; IFN-g, 20 ng/mL; lane 2), TSA (300 nmol/L; lane 3), or IFNs and TSA together for 24 h were analyzed by immunoblotting using antibodies specific to the indicated proteins.

Article Snippet: Mammalian expression vector (pCMV6-JAK1) to express human JAK1 protein was purchased from OriGene Technologies, Inc. (Rockville, MD).

Techniques: Expressing, Control, Western Blot, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: Loss of Janus Associated Kinase 1 Alters Urothelial Cell Function and Facilitates the Development of Bladder Cancer

doi: 10.3389/fimmu.2019.02065

Figure Lengend Snippet: STAT1 phosphorylation and expression of IRF1 mRNA is impaired in JAK1-deficient hTERT urothelial cells. (A) JAK1 protein (InterPro P23458) structure showing core domains, TCGA BLCA variants and their predicted impact. Protein domains: FERM (4.1 protein, ezrin, radizin, moesin domain), PHD (pleckstrin homology-like domain), SH2 (Src homology 2), S-TY-PK (unknown specificity serine-threonine/tyrosine protein kinase), SY-PK (serine-tyrosine protein kinase). Variants: non-sense (red cross), missense (yellow inverted triangle), synonymous (green triangle), frameshift (purple square). Variant effects: S (SIFT score < 0.05), P (Poly-Phen score > 0.908), p (Poly-Phen score > 0.446 & ≤ 0.908), H (HMMvar score > 2). A single 5′ UTR modifier mutation not shown. (B,C) Analysis of JAK/STAT signaling by flow cytometry in untransduced (Unt), Sc and KD hTERT immortalized urothelial cell lines after stimulation with IFNγ (1 ng/ml) for 24 h. Data (A) is from a representative experiment, data (B) is from three independent experiments. Two-tailed Mann Whitney test. (D) RTqPCR analysis of IRF1 mRNA expression from KD and Sc hTERT immortalized urothelial cell lines after stimulation with IFNγ (1 ng/ml). Data is from four independent experiments. Two-tailed Mann Whitney test. * P < 0.05 Error bars represent the SE. (E,F) Flow cytometry analysis of IFNGR expression in KD and Sc hTERT immortalized urothelial cell lines following stimulation with IFNγ (5 ng/ml) for 2 days. The graph shows mean values ± SD. Data is from three independent experiments. One-way ANOVA with Tukey's Multiple Comparison Test.

Article Snippet: Mutations and their effects were presented against the JAK1 protein sequence (InterPro P23458).

Techniques: Phospho-proteomics, Expressing, Variant Assay, Mutagenesis, Flow Cytometry, Two Tailed Test, MANN-WHITNEY, Comparison

Journal: Frontiers in Immunology

Article Title: Loss of Janus Associated Kinase 1 Alters Urothelial Cell Function and Facilitates the Development of Bladder Cancer

doi: 10.3389/fimmu.2019.02065

Figure Lengend Snippet: JAK1-deficient hTERT urothelial cells demonstrate preserved population growth and reduced apoptosis in response to IFNγ. (A) KD and Sc hTERT immortalized urothelial cell lines were cultured with addition of Alamar Blue (AB) dye and stimulated with IFNγ (5 ng/ml) for the given time points. The capacity of viable cells to reduce AB dye was used as a proxy for cell number. Data (A) is from five independent experiments. (B–E) KD and Sc hTERT urothelial cell lines were stimulated with the given concentrations of IFNγ for 5 days. Percentage of early and late apoptosis was quantified with Annexin V/PI apoptosis detection kit by flow cytometry. Data (B–D) are from five independent experiments, data (E) is from a representative experiment. One-way ANOVA with Tukey's Multiple Comparison post-test. * P < 0.05. Error bars represent the SE.

Article Snippet: Mutations and their effects were presented against the JAK1 protein sequence (InterPro P23458).

Techniques: Cell Culture, Flow Cytometry, Comparison

Journal: Frontiers in Immunology

Article Title: Loss of Janus Associated Kinase 1 Alters Urothelial Cell Function and Facilitates the Development of Bladder Cancer

doi: 10.3389/fimmu.2019.02065

Figure Lengend Snippet: JAK1-deficient hTERT urothelial cells showed reduced lymphocyte-mediated killing in response to IFNγ. JAK1-deficient and Sc hTERT immortalized urothelial cell lines were pretreated or not with IFNγ and cultured overnight with 25 U/ml IL-2 and monocyte-depleted PBMCs (50:1). (A,B) Necrosis induction in JAK1-deficient and Sc hTERT immortalized urothelial cell lines. One-way ANOVA with Dunn's multiple comparisons post-test. (C) Relative change of necrosis compared to untreated. Two-tailed Mann Whitney U -test. Data is from five independent experiments. * P < 0.05. Error bars represent the SE.

Article Snippet: Mutations and their effects were presented against the JAK1 protein sequence (InterPro P23458).

Techniques: Cell Culture, Two Tailed Test, MANN-WHITNEY