human lambda light chain  (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