Journal: Experimental and Therapeutic Medicine

Article Title: A rare dermatological manifestation of follicular spicules in a patient with multiple myeloma and end-stage renal disease on hemodialysis: A case report

doi: 10.3892/etm.2025.12930

Figure Lengend Snippet: SPE and immunofixation confirming monoclonal gammopathy. (A) SPE demonstrating an M-spike in the γ region. (B) Immunofixation electrophoresis revealing a monoclonal IgA λ pattern, confirming the presence of IgA-λ type multiple myeloma. (C) Immunofixation electrophoresis further confirming the monoclonal IgA λ pattern. AU, arbitrary units; SPE, serum protein electrophoresis. ELP, electrophoresis reference; G, IgG; A, IgA; M, IgM; K, κ light chain; L, λ light chain; D, IgD; E, IgE.

Article Snippet: Primary antibodies were incubated overnight at 4 ̊C, including: CD138 (cat. no. ab34164; dilution 1:100; Abcam), CD56 (cat. no. ab75813; dilution 1:200; Abcam), κ light chain (cat. no. ZM-0317; dilution 1:150; OriGene Technologies, Inc.) and λ light chain (cat. no. ZM-0318; dilution 1:150; Origene Technologies, Inc.).

Techniques: Electrophoresis, Protein Electrophoresis

Journal: Experimental and Therapeutic Medicine

Article Title: A rare dermatological manifestation of follicular spicules in a patient with multiple myeloma and end-stage renal disease on hemodialysis: A case report

doi: 10.3892/etm.2025.12930

Figure Lengend Snippet: Immunohistochemical characterization of bone marrow biopsy. (A) Congo red staining showed amyloid deposits with birefringence under polarized light, confirming amyloidosis. (B) Scattered CD3 + T cells. (C) CD20 + B cells. (D) Diffuse CD56 positivity indicating natural killer cell involvement. (E) Diffuse CD138 staining marking plasma cell infiltration. (F) Negative κ light chain staining. (G) Diffuse positive staining of λ light chains, confirming monoclonality. (H) Multiple myeloma oncogene 1-positivity indicated plasma cell differentiation. (I) Hematoxylin and eosin staining revealed diffuse infiltration by atypical lymphoid/plasma cells. (J) Positive staining for amyloid deposits in bone marrow biopsy. All images were captured under a light microscope at x400 magnification.

Article Snippet: Primary antibodies were incubated overnight at 4 ̊C, including: CD138 (cat. no. ab34164; dilution 1:100; Abcam), CD56 (cat. no. ab75813; dilution 1:200; Abcam), κ light chain (cat. no. ZM-0317; dilution 1:150; OriGene Technologies, Inc.) and λ light chain (cat. no. ZM-0318; dilution 1:150; Origene Technologies, Inc.).

Techniques: Immunohistochemical staining, Staining, Clinical Proteomics, Cell Differentiation, Light Microscopy

Journal: Experimental and Therapeutic Medicine

Article Title: A rare dermatological manifestation of follicular spicules in a patient with multiple myeloma and end-stage renal disease on hemodialysis: A case report

doi: 10.3892/etm.2025.12930

Figure Lengend Snippet: Flow cytometric immunophenotyping of bone marrow cells from a patient with PCM. (A) Lymphocyte analysis showing a decreased CD4/CD8 ratio. (B and C) Quantification of CD57⁺ LGLs, demonstrating a normal proportion of this subset. (D) Marked expansion of plasma cells detected within the bone marrow aspirate. (E) Dual-parameter plot of CD38 and CD45 expression revealing a distinct CD38 bright+ /CD45 dim+ population, accounting for 33.19% of total nucleated cells, consistent with malignant plasma cell infiltration. (F) Intracellular immunoglobulin light chain staining within the CD38 bright+ population indicating prominent λ light chain restriction (97.37%) with minimal κ expression (0.03%), confirming clonal proliferation and supporting the diagnosis of PCM. LGLs, large granular lymphocytes; NC, nucleated cell; PCM, plasma cell myeloma; A, allophycocyanin-Alexa Fluor 700; APC, allophycocyanin; CD38bri + , CD38 bright positive; DNT, double negative T cells; DPT, dual-parameter plot; ECD, energy-coupled dye; H, height parameter; KO525, Krome Orange 525; Lym, lymphocytes; NK, natural killer cells; PC5.5, phycoerythrin-cyanine 5.5; PE, phycoerythrin; SSC, side scatter.

Article Snippet: Primary antibodies were incubated overnight at 4 ̊C, including: CD138 (cat. no. ab34164; dilution 1:100; Abcam), CD56 (cat. no. ab75813; dilution 1:200; Abcam), κ light chain (cat. no. ZM-0317; dilution 1:150; OriGene Technologies, Inc.) and λ light chain (cat. no. ZM-0318; dilution 1:150; Origene Technologies, Inc.).

Techniques: Clinical Proteomics, Expressing, Staining, Biomarker Discovery

Journal: Oncotarget

Article Title: Mechanisms for autophagy modulation by isoprenoid biosynthetic pathway inhibitors in multiple myeloma cells

doi:

Figure Lengend Snippet: A . RPMI-8226, ALMC-2, MM.1S, and U266 cells were incubated for 48 hours in the presence or absence of lovastatin ( Lov , 2-50 μM) or DGBP (2-50 μM). Immunoblot analysis of p62, Atg3 and β-tubulin (loading control) is shown. The relative intensity ( Rel Int) values underneath the p62 and Atg3 blots represent relative levels as determined by densitometric analysis of Atg3 or p62 (relative to tubulin) of treated cells compared to control cells. The gels are representative of 3-4 independent experiments. B . ALMC-2 cells were incubated for 48 hours in the presence of 10 μM lovastatin ( Lov ) and/or mevalonate ( Mev , 5 mM), FPP (10 μM), or GGPP (10 μM) for 48 hrs. Densitometric analysis of p62 or Atg3 levels (normalized to β-tubulin levels) for the combination treatments normalized to untreated cells (control) is shown. Data are displayed as average +/− standard deviation (n=3 independent experiments). * denotes p-value <0.05 from two-sided t-testing. C . RPMI-8226 and ALMC-2 cells were incubated for 48 hours in the presence or absence of 10 μM lovastatin ( Lov ), 10 μM DGBP, 10 μM GGTI-2133 ( GGTI ), 10 mM 3-PEHPC ( 3P ), or the combination of 10 μM GGTI-2133 and 10 mM 3-PEHPC ( GGTI + 3P ). Immunoblot analysis of p62, Atg3 and β-tubulin (loading control) is shown. The gels are representative of 3-4 independent experiments. D . RPMI-8226 and ALMC-2 cells were incubated for 48 hours in the presence or absence of 10 μM lovastatin ( Lov ), 10 μM DGBP, 10 mM 3-PEHPC ( 3P ), or 5 nM bafilomycin A1 ( Baf ) with or without protease inhibitors (10 μg/mL E64d and pepstatin A, prot ). Immunoblot analysis of LC3 and β-tubulin (loading control) is shown. The gels are representative of 3 independent experiments. E . Intracellular lambda light chain levels were measured via ELISA. Data are expressed as a percentage of control (mean +/− standard deviation of 3 independent experiments). Two-sided t-testing was performed to compare the IBP inhibitor alone vs. in combination with the protease inhibitors. No significant differences between the treatment groups were observed.

Article Snippet: A human lambda light chain kit (E80-116, Bethyl Laboratories) was used to quantify intracellular monoclonal protein levels.

Techniques: Incubation, Western Blot, Control, Standard Deviation, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Mechanisms for autophagy modulation by isoprenoid biosynthetic pathway inhibitors in multiple myeloma cells

doi:

Figure Lengend Snippet: RPMI-8226 cells were incubated with 2 nM bafilomycinA1, 10 μM lovastatin, or 5 mM 3-PEHPC for 48 hours. Staining was performed as described in Materials and Methods, using antibodies directed against lambda light chain (red) and PDI as an ER marker (green) in A . and RCAS1 as a Golgi marker (green) in B . DAPI was used for nuclear staining (blue). Colocalization was determined using the RGB plot profile tool in ImageJ and indicated by similarity of the patterns of red and green peaks. Scale bar 10μm.

Article Snippet: A human lambda light chain kit (E80-116, Bethyl Laboratories) was used to quantify intracellular monoclonal protein levels.

Techniques: Incubation, Staining, Marker

Journal: Oncotarget

Article Title: Mechanisms for autophagy modulation by isoprenoid biosynthetic pathway inhibitors in multiple myeloma cells

doi:

Figure Lengend Snippet: A . RPMI-8226 cells were incubated with 2 nM bafilomycin A1 ( Baf ), 10 μM lovastatin ( Lov ), or 5 mM 3-PEHPC ( 3P ) for 48 h or 10 nM bortezomib for 24 h. Staining was performed as described in Materials and Methods, using Proteostat (Enzo Life Sciences, Inc.) (green) to visualize aggresomes and an antibody directed against lambda light chain (red). DAPI was used for nuclear staining (blue). Colocalization was determined using the RGB plot profile tool in ImageJ. Scale bar 10 μm. B . The average number of aggresomes per cell was quantified using the GFP-LC3 macro for ImageJ (mean ± SEM). **** denotes P <0.0001 from unpaired two-tailed t- test comparing treated with control cells.

Article Snippet: A human lambda light chain kit (E80-116, Bethyl Laboratories) was used to quantify intracellular monoclonal protein levels.

Techniques: Incubation, Staining, Two Tailed Test, Control

Journal: Oncotarget

Article Title: Mechanisms for autophagy modulation by isoprenoid biosynthetic pathway inhibitors in multiple myeloma cells

doi:

Figure Lengend Snippet: U266 cells were treated with 2 nM bafilomycin A1 ( Baf ), 10 μM lovastatin ( Lov ) or 10 mM 3-PEHPC for 48 hours before processing for staining with LC3. A . The average number of LC3 puncta per cell and B . the average size of each punctate structure in pixels was quantified using the GFP-LC3 macro for ImageJ (mean ± SEM) (n>175). The **** denotes p <0.0001 from unpaired two-tailed t- test comparing treated with control cells. C . Representative confocal maximum projections are shown for each treatment with LC3 in green and DAPI staining the nucleus in blue. D . RPMI-8226 cells were incubated with 2 nM bafilomycin A1, 10μM lovastatin, 5mM 3-PEHPC or a co-treatment with bafilomycin A1 ( Baf ) and lovastatin ( Lov ) for 48 hours. Staining was performed as described in Materials and Methods, using CytoID (Enzo Life Sciences, Inc., Farmingdale, NY) to visualize autophagosomes and an antibody directed against lambda light chain. DAPI was used for nuclear staining. Colocalization was determined using the RGB plot profile tool in ImageJ. Scale bar 10μm.

Article Snippet: A human lambda light chain kit (E80-116, Bethyl Laboratories) was used to quantify intracellular monoclonal protein levels.

Techniques: Staining, Two Tailed Test, Control, Incubation

Journal: Blood Cancer Journal

Article Title: Downregulation of PA28α induces proteasome remodeling and results in resistance to proteasome inhibitors in multiple myeloma

doi: 10.1038/s41408-020-00393-0

Figure Lengend Snippet: a Western blot analysis of ubiquitinated proteins in total protein lysates extracted from LP1 and RPMI8226 PA28α knockdown stable cells. β-actin as a loading control. b OPP pulse-chase assay of proteasome degradation and protein synthesis in LP1 PA28α knockdown stable cells. Western blot analysis of immunoglobulin lambda light chain (ƛ IgL) ( c ), eIF2α, p62/SQSTM1, and LC3B ( d ) in LP1 and RPMI8226 PA28α knockdown stable cells, β-actin as a loading control. ** P < 0.01, Student’s t test. e Working model of PA28α knockdown in MM.

Article Snippet: Antibodies used were as follows: PA28α (Cell Signaling), PSMA2 (Cell Signaling), S5a (Cell signaling), PA28β (Cell Signaling), Phospho-eIF2α (Ser51) (Cell signaling), eIF2α (Cell signaling), α-tubulin (Genetex), PA28γ (Genetex), PSMB5 (Genetex), PSMB6 (Enzo life science), PSMB7 (Genetex), PSMB8 (Genetex), PSMB9 (R&D systems), PSMB10 (R&D systems), Rpt5 (Enzo life science), Ubiquitin (Cell Signaling), β-actin (Santa Cruz Technology), TCF11/NRF1 (Cell Signaling), LC3B (Cell Signaling), p62/SQSTM1 (MBL International) human Ig lambda light chain (R&D systems), actin (Sigma). pLKO.1 empty vector, shRNA vector targeting human PA28α, NRF1 siRNA (ON-TARGETplus SMARTpool), PA28α siRNA (Accell SMARTpool), and control siRNA were purchased from Dharmacon.

Techniques: Western Blot, Knockdown, Control, Pulse Chase