Journal: Scientific Reports

Article Title: Identified endoplasmic reticulum stress-related molecular cluster and immune characterization in endometriosis

doi: 10.1038/s41598-025-22400-9

Figure Lengend Snippet: qRT-PCR and IHC validation the hub gene expression level. ( a ) Hub ERS genes mRNA expression of normal, eutopic and ectopic endometrium. ( b ) Immunostaining of EPAS1, F8, VCAM1, VWF protein expression. ( c ) Semi-quantitative analysis of IHC results. qRT-PCR, real-time quantitative reverse transcription; IHC, immunohistochemistry. * p < 0.05, ** p < 0.01, ∗ ∗ ∗ p < 0.001, **** p < 0.0001.

Article Snippet: Briefly, the sections were stained with the primary antibody against F8 (1:500, affinity, USA), VCAM1(1:200, Proteintech, Wuhan, China), VWF (1:400, Proteintech, Wuhan, China), and EPAS1(1:50, Proteintech, Wuhan, China) and then scanned with a digital scanner.

Techniques: Quantitative RT-PCR, Biomarker Discovery, Gene Expression, Expressing, Immunostaining, Reverse Transcription, Immunohistochemistry

Journal: Science Advances

Article Title: PHF8-GLUL axis in lipid deposition and tumor growth of clear cell renal cell carcinoma

doi: 10.1126/sciadv.adf3566

Figure Lengend Snippet: ( A ) Representative images and quantification data of lipid droplets stained with BODIPY 493/503 (green) in 786-O with or without VHL reexpression (Re VHL ). Nuclei were stained with DAPI (blue; scale bar, 50 μm). EV, empty vector. ( B ) Real-time quantitative polymerase chain reaction (RT-qPCR) analysis of relative mRNA expressions of seven JmjC-containing epigenetic regulators in 786-O with or without Re VHL . ( C ) Flow cytometric analysis of lipid droplets in 786-O cells transfected with siRNAs targeting indicated genes using BODIPY 493/503. ( D ) Representative images and quantification data of lipid droplets stained with ORO (red) in indicated 786-O cells. Nuclei stained with Mayer’s hematoxylin (scale bar, 100 μm in 400×). ( E ) WB analysis of the indicated proteins in 786-O cells with or without Re VHL . ( F ) Chromatin immunoprecipitation (ChIP)–qPCR analysis of hypoxia-inducible factor 1α (HIF1α) binding on the promoter regions of PHF8 and GLUT1 . IgG, immunoglobulin G. ( G ) ChIP-qPCR analysis of HIF2α binding on the promoter regions of PHF8 and EPO . ( H ) PHF8 promoter–driven luciferase reporters and luciferase activity (LUC) in 786-O cells with overexpressed HIF1α and HIF2α. ( I and J ) RT-qPCR analysis of relative mRNA expressions of indicated genes in 786-O Re VHL cells with or without overexpression of nondegradable mut HIF1A (P402A/P564A) (I) or mut HIF2A (P405A/P531A) (J). ( K ) Representative images and quantification data of lipid droplets stained with ORO (red) in indicated 786-O cells. Nuclei stained with Mayer’s hematoxylin (scale bar, 100 μm in 400×). In (A), (B), (D), and (F) to (K), data are presented as means ± SD from at least three independent experiments or biological replicates. Two-tailed Student’s t test was used for (A), (B), and (F) to (H), and ordinary one-way analysis of variance (ANOVA) was used for (D) and (I) to (K). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: MYC- HIF2A was obtained from OriGene (RC208604).

Techniques: Staining, Plasmid Preparation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Chromatin Immunoprecipitation, ChIP-qPCR, Binding Assay, Luciferase, Activity Assay, Over Expression, Two Tailed Test

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: 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, B Cell viability assay of MOP-1 (top panel, mean ± s.e.m, n = 3), MOP-2 (second panel, mean ± s.e.m, n = 3), ZrMOP (third panel, mean ± s.e.m, n = 6) and ligand (bottom panel, mean ± s.e.m, n = 5). Schematics (left panel) were created with Diamond software. A MOPs treated mESC (J1 cell line) exhibited flat morphology compared with the mESC cultured in LIF addition medium (denoted as LIF mESC) and mESC cultured without LIF (denoted as Ctrl mESC), which grew in colonies and expressed positive alkaline phosphatase (ALP) staining ( B ). Scale bar, 100 μm. C Representative images of ALP staining (left panel) and quantification area of LIF and MOP-1 mESC colonies (right panel) (mean ± s.e.m, n = 3). Scale bars, 40 μm). D The cell cycle distribution of MOP-1 mESC was evaluated by flow cytometry. The percentage of G0/G1, S, G2/M phases of the cell cycle (mean ± s.e.m, n = 3). E Plots of FITC-annexin V/PI flow cytometry of mESC cultured with or without MOP-1 for 48 h. F Representative immunoblot images (left panel) and quantification (right panel) of p-SHP-2 in MOP-1 mESC under different concentrations (mean ± s.e.m, n = 3). G Representative immunoblot images of STAT3 in LIF mESC and MOP-1 mESC at different withdrawal times (mean ± s.e.m, n = 4). Data in ( C ) and ( D ) are analyzed by a two-tailed unpaired t -test. Data in ( F ) are analyzed by one-way ANOVA. # P < 0.05, 0 μM MOP-1 mESC vs. LIF mESC; * P < 0.05, 2 μM and 4 μM MOP-1 mESC vs. 0 μM MOP-1 mESC; no statistical significance (NS), LIF mESC vs. 2 μM and 4 μM MOP-1 mESC, MOP-1 mESC vs. LIF mESC or Ctrl mESC.

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: Viability Assay, Software, Cell Culture, Staining, Flow Cytometry, Western Blot, Two Tailed Test

Journal: Clinical and Experimental Medicine

Article Title: Therapeutic potential of third-generation chimeric antigen receptor T cells targeting B cell maturation antigen for treating multiple myeloma

doi: 10.1007/s10238-024-01347-7

Figure Lengend Snippet: Lentiviral constructs of anti-BCMA-CARs and protein expression in Lenti-X™ HEK293T cells. A Schematic representation shows second-generation anti-BCMA-CAR (anti-BCMA-CAR2) and third-generation anti-BCMA-CAR (anti-BCMA-CAR3) lentiviral constructs containing fully human anti-BCMA scFv, c-Myc tag, hinge region, transmembrane (TM) domain, co-stimulatory domain(s), and CD3ζ. B Representative histogram demonstrates the expression of anti-BCMA-CARs on the surface of Lenti-X™ HEK293T cells analyzed by flow cytometry using by anti-cMyc-FITC antibody, and C the data were summarized as bar graphs. D Immunoblot analysis of cell lysates of Lenti-X™ HEK293T cells using anti-CD3ζ antibody shows specific bands of anti-BCMA-CAR2 and anti-BCMA-CAR3 at 62 and 66 kDa, respectively, with a loading control of 37 kDa GAPDH. E The densitometry data of anti-BCMA-CAR protein bands relative to GAPDH bands on SDS-PAGE analyzed using ImageJ software were summarized as bar graphs. The data obtained from 3 independent experiments were expressed as mean ± standard error of the mean (SEM) (N = 3). Statistical significance was determined using one-way analysis of variance (ANOVA) with Tukey’s post-hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001)

Article Snippet: The fully human anti-BCMA scFv codon-optimized gene was sub-cloned into a self-inactivating (SIN) lentiviral vector (pCDH.EF1α.SIN.WPRE, RRID: Addgene_71708) containing expression cassettes encoding CD8 short hinge, CD8 transmembrane domain, and the 4-1BB/CD3ζ signaling domain (anti-BCMA-CAR2) or CD8 short hinge, CD28 transmembrane domain, and the CD28/4-1BB/CD3ζ signaling domain (anti-BCMA-CAR3) (Fig. A).

Techniques: Construct, Expressing, Flow Cytometry, Western Blot, Control, SDS Page, Software