intracellular lambda light chain levels  (Bethyl)

Journal: Nature Communications

Article Title: EZH2 serves as a viable therapeutic target for myeloma-induced osteolytic bone destruction

doi: 10.1038/s41467-025-56506-5

Figure Lengend Snippet: 6–7 weeks old female NSG mice were intrafemorally injected with myeloma cell lines RPMI8226 [nontargeted shRNA (sh Ctrl ), and EZH2 shRNA (sh EZH2 )] or MM.1S [control vector ( Vec ), and EZH2 cDNA ( EZH2 )]. The mice not receiving myeloma cells (No MM) served as controls. After six weeks of cells injection, shown are the representative x-ray images of lytic lesions ( a ) (n = 5 mice/group). Yellow arrow, bone lesion. Scale bar, 1 mm. Another batch of NSG mice were intrafemorally injected with the same combination of myeloma cell lines. After four weeks of injections, shown are the representative images of bioluminescent signals ( b ) and microcomputed tomography images ( c ), concentrations of PINP ( d ) or CTX-1 ( e ) in mouse sera, percentages of BV/TV ( f ), ES/BS ( g ), Oc.S/BS ( h ), OS/BS ( i ), and Ob.S/BS ( j ). Scale bar, 1 mm. k , l Bone formation rate (BFR/BS) was measured by calcein injection, and the bone sections were imaged and analyzed. Shown are representative images and summarized data of bone formation in mouse femurs (n = 5 mice/group). Scale bar: 20 μm. m Schematic diagram showing primary patient-derived mouse model generation and analyzes performed. n ELISA analysis shown the concentrations of human λ chains in mouse serum. o After the mice were euthanized, myeloma cells were isolated from the bone marrow of PDX mice using anti-CD138 antibody-coated magnetic beads. EZH2 mRNA levels were compared between P1 and P2 groups (n = 5 mice/group). p Shown are the representative microcomputed tomography images in PDX mouse model. Scale bar, 0.5 mm. Shown are the percentages of BV/TV ( q ), ES/BS ( r ), and OS/BS ( s ) in PDX mouse model. The P1 and P2 cells used here are from the same source as those in Fig. . d – k , q – s : p values were determined using one-way ANOVA with Tukey’s test; ( n , o ): p values were determined by unpaired two-tailed t test. Data are means ± SD. Source data are provided as a file.

Article Snippet: Human λ chains ELISA Kits were purchased from Bethyl Laboratories (#E88-116).

Techniques: Injection, shRNA, Control, Plasmid Preparation, Tomography, Derivative Assay, Enzyme-linked Immunosorbent Assay, Isolation, Magnetic Beads, Two Tailed Test

Journal: Nature Communications

Article Title: EZH2 serves as a viable therapeutic target for myeloma-induced osteolytic bone destruction

doi: 10.1038/s41467-025-56506-5

Figure Lengend Snippet: a A published GEO dataset (GSE103567) analysis shows the expression of cytokines in RPMI8226 cell treated with or without GSK343 (4 μM). b , c The expression of CCL5 , IL16 , CSF1 and DKK1 in RPMI8226 (sh Ctrl , sh EZH2 ) or MM.1S ( Vec , EZH2 ) cells (n = 3 biological replicates). d – g ELISA analysis shows the secretion of CCL5, IL16, CSF1 and DKK1 in RPMI8226 (sh Ctrl , sh EZH2 ) or MM.1S ( Vec , EZH2 ) (n = 3 biological replicates). Correlation coefficient of the levels of CCL5 ( h ), IL16 ( i ), CSF1 ( j ) or DKK1 ( k ) and levels of EZH2 (n = 30). The cells used here are from the same source as those in Fig. . The correlation was evaluated using Pearson coefficient with two-tailed p value. r, correlation coefficient. l The numbers of multinuclear TRAP + cells among precursors of osteoclasts cultured with MM.1S cells ( Vec or EZH2 ) in the presence of blocking antibodies against CCL5, IL16, CSF1 or DKK1 (1 μg/ml) (n = 3 biological replicates). m The Alizarin red S staining of MSCs cultured in osteoblast medium with MM.1S cells ( Vec or EZH2 ) cells in the presence of antibodies against CCL5, IL16, CSF1 or DKK1 (1 μg/ml) (n = 3 biological replicates). n The numbers of multinuclear TRAP + cells among precursors of osteoclasts cultured with RPMI8226 cells (sh Ctrl or sh EZH2 ) in the presence of CCL5 (20 ng/ml), IL16 (2 ng/ml), CSF1 (10 ng/ml) or DKK1 (20 ng/ml) (n = 3 biological replicates). o The Alizarin red S staining of MSCs cultured in osteoblast medium with RPMI8226 cells (sh Ctrl or sh EZH2 ) cells in the presence of CCL5 (20 ng/ml), IL16 (2 ng/ml), CSF1 (10 ng/ml) or DKK1 (20 ng/ml) (n = 3 biological replicates). Data are means ± SD. b – g : p values were determined by unpaired two-tailed t test; l – o : p values were determined using one-way ANOVA with Tukey’s test. Source data are provided as a file.

Article Snippet: Human λ chains ELISA Kits were purchased from Bethyl Laboratories (#E88-116).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Cell Culture, Blocking Assay, Staining

Journal: Nature Communications

Article Title: EZH2 serves as a viable therapeutic target for myeloma-induced osteolytic bone destruction

doi: 10.1038/s41467-025-56506-5

Figure Lengend Snippet: a , b We pre-treated myeloma cells (RPMI8226 or MM.1S) with DZNep (1 μM) or GSK343 (4 μM) and collected conditional medium. Shown are the formation of TRAP + cells from preOCs and Alizarin red-S staining for osteoblast differentiation from MSCs, cultured with different myeloma cells conditional medium (30% volume ratio). Addition of PBS or no addition of conditional medium served as a control (n = 4 biological replicates). 6–7 weeks old female NSG mice were intravenously injected with luciferase-labeled RPMI8226 cells, and intraperitoneal administration of DZNep or GSK343 (100 μg/kg body weight) three times per week for a duration of two weeks, beginning two weeks after cell injection. c Representative images and summarized data of bioluminescent signals in NSG mice (n = 5 mice/group). d Shown are the representative microcomputed tomography images. NSG mice left femurs were extracted, fixed, TRAP- or toluidine blue-stained, and analyzed (n = 5 mice/group). Scale bar: 0.5 mm. e Representative images of TRAP- or toluidine blue-stained femurs of mice (n = 5 mice/group). Scale bar: 50 μm. Black arrow, osteoblast. Shown are the percentages of BV/TV ( f ), ES/BS ( g ), Oc.S/BS ( h ), OS/BS ( i ), Ob.S/BS ( j ), and human λ chain levels ( k ) (n = 5 mice/group). l ELISA analysis shown the concentrations of human CCL5, IL16, CSF1 and DKK1 in mouse serum (n = 5 mice/group). m After treatment, myeloma cells were also isolated from the right femurs of NSG mice using anti-CD138 antibody-coated magnetic beads. EMP1 mRNA levels were compared in isolated myeloma cells treated with or without EZH2 inhibitors (n = 5 mice/group). n Depiction of EZH2-mediated signaling pathways in myeloma cells and adipocytes. MM, multiple myeloma. Data are means ± SD. All p values were determined using one-way ANOVA with Tukey’s test. Source data are provided as a file.

Article Snippet: Human λ chains ELISA Kits were purchased from Bethyl Laboratories (#E88-116).

Techniques: Staining, Cell Culture, Control, Injection, Luciferase, Labeling, Tomography, Enzyme-linked Immunosorbent Assay, Isolation, Magnetic Beads, Protein-Protein interactions

Journal: JCI Insight

Article Title: An effective cell-penetrating antibody delivery platform

doi: 10.1172/jci.insight.127474

Figure Lengend Snippet: (A) Tumor growth kinetics when treated with indicated modified antibodies (n = 8). SD shown. Two-way ANOVA (Tukey’s multiple-comparisons test) was used: ***P < 0.001. (B) ELISA on pY(418)-Src recognition by anti-pY(418)Src antibody upon modification performed twice in triplicates. (C) Representative Western blot assessing gene expression in tissue homogenates isolated from tumors. (D) Tumor tissue integrity, tumor accumulation of modified antibodies and CD31+ tumor vasculature, and pY(418)-Src levels shown by confocal microscopy (left) with quantification (right). (E) Representative RT-PCR on Src kinase target gene mRNA expression in tumors treated as indicated, performed in triplicates. (B, D, and E) SD shown. Unpaired Student’s t test: **P < 0.01, and ***P < 0.001.

Article Snippet: Human IgG λ chain was detected using the human IgG λ light chain ELISA kit according to the manufacturer’s instructions (MyBioSource.com).

Techniques: Modification, Enzyme-linked Immunosorbent Assay, Western Blot, Gene Expression, Isolation, Confocal Microscopy, Reverse Transcription Polymerase Chain Reaction, Expressing