Journal: Neoplasia (New York, N.Y.)

Article Title: SAMSN1 Is a Tumor Suppressor Gene in Multiple Myeloma 1 2

doi: 10.1016/j.neo.2014.07.002

Figure Lengend Snippet: SAMSN1 expression is reduced in CD138 + PCs of patients with MM and HMCL. (A) SAMSN1 expression (as determined by real-time PCR) is significantly reduced in the BMs of patients with MM ( n = 34) compared with patients with MGUS ( n = 9) and healthy age-matched controls ( n = 5; * P < .05, ** P < .001, one-way ANOVA with Tukey’s multiple comparison test). (B) SAMSN1 expression in CD138 + MACS isolated PCs from patients with MM negatively correlates with BM PC burden ( n = 10, r 2 = 0.6147, P = .0043). (C) In silico analysis of published microarray data. CD138 + PCs were isolated by MACS from 414 patients with MM, 44 patients with MGUS, and 22 age-matched controls. RNA was extracted and analyzed using the Affymetrix U133Plus2.0 microarray platform (GEO Accession Nos GSE4581 and GSE5900). Expression of SAMSN1 is significantly reduced in PCs of patients with MM compared to those of patients with MGUS and normal controls. P < 0.0001, one-way ANOVA with Tukey’s multiple comparison test. (D) Total RNA was extracted from six HMCLs and reverse transcribed. The levels of SAMSN1 expression were assessed by real-time PCR.

Article Snippet: PCs were isolated from flushed long bones and identified using rat anti-mouse CD138 (R&D Systems, Minneapolis, MN) followed by PE-conjugated goat anti-rat IgG.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Comparison, Isolation, In Silico, Microarray, Reverse Transcription

Journal: Nature Communications

Article Title: GWAS for Interleukin-1β levels in gingival crevicular fluid identifies IL37 variants in periodontal inflammation

doi: 10.1038/s41467-018-05940-9

Figure Lengend Snippet: Expression and localization of IL-37 in human gingival tissue. a The sketch of five IL-37 isoforms. b PCR products amplified with specific primer pairs to different isoforms of IL-37 cDNA. c The relative mRNA levels of different isoforms of IL-37 in human gingival tissues from sites with periodontitis were quantitated using real time PCR with isoform-specific primers. d The relative mRNA levels of IL-37b (isoform1) in gingival tissues between health and periodontitis ( n ≥ 5/group). Data show the mean ± SD. *** p < 0.0001 (unpaired two-tailed Student’s t -test). e The representative level of IL-37 positive staining in epithelial and connective tissue of human gingival tissue from healthy and periodontitis patient using IHC. Scale bar is 200 µm. f The localization of IL-37b and CD138 were determined in serial slides of gingival tissue from periodontitis patient. The localization of IL-37 in human gingival tissue was mainly associated with epithelial and infiltrated plasma cells. The black circles with red number indicate representative region of IL-37 positive staining. A , B represent the low magnification of IL-37 and CD138 staining in human gingival tissue with Scale bar 200 µm. C – F represent the high magnification of two immune cells infiltrated regions in the connective tissue with Scale bar 20 µm. g , h represent the high magnification of epithelial cells layer with Scale bar 20 µm. g IL-37b and CD138 were co-localized in human gingival tissue of periodontitis patient by immunofluorescence using confocal microscope. The results represent CD138 (green), IL-37 (red), DAPI (blue), and bright field (white). The scale bar is 20 µm. Data shown here are representative of three independent experiments

Article Snippet: The slides were stained with anti-IL-37 antibody with 1:1000 dilution (rabbit anti-human, ThermoFisher SCIENTIFIC, PA5-30527), anti-CD138 with 1:150 dilution (goat anti-human CD138, R&D SYSTEMS, AF2780) antibody.

Techniques: Expressing, Amplification, Real-time Polymerase Chain Reaction, Two Tailed Test, Staining, Clinical Proteomics, Immunofluorescence, Microscopy

Journal: bioRxiv

Article Title: Anti-NMDA receptor encephalitis in mice induced by active immunization with conformationally-stabilized holoreceptors

doi: 10.1101/467902

Figure Lengend Snippet: (A) Robust expression of the panleukocyte marker, CD45R in a proteoliposome treated-mouse (right). Insets show labeling of individual CD45R + cells (white) from the indicated region of the hippocampus. Coronal sections from control mice had few if any CD45R + cells (left). Scale bar: 1000μm (inset 100μm). (B) Control mice (left) had sparse or absent labeling for a battery of immune cell markers (CD8 + , CD4 + , CD20 + , CD138 + , Gal3 + ). Proteoliposome treated-mice showed significant increases in immune cell infiltrates as indicated with the insets from the hippocampal region; [Scale bar: 500μm (inset 200μm). (C) Left: CD45R + density (cells/μm 3 ) in striatum (Str), cortex (Ctx), amygdala (Amyg), hippocampus (Hipp) and thalamus (Thal) in proteoliposome-treated (black) and control mice (gray). Right: CD8 + , CD4 + , CD20 + , CD138 + , Galectin3 + cell densities (cells/μm 3 ) in the hippocampus of proteoliposome treated (black) and control mice (gray). Only Gal3 + cells were of sufficient density to be seen on the histogram in control mice.

Article Snippet: The following primary/secondary antibody pairings were used to label floating brain section: CD45R (BD Biosciences #550539, 1:25; Invitrogen #A21208, AlexaFluor488), CD4 (BD Biosciences #550280, 1:25; Invitrogen #A21208, AlexaFluor488), CD8 (BD Biosciences #550281, 1:25; Invitrogen #A21208, AlexaFluor488), CD20 (Santa Cruz Biotechnology #7735, 1:50; Invitrogen #A21447, AlexaFluor647), CD138 (R&D Systems #AF3190, 1:100; Invitrogen #A11057, AlexaFluor568), Galectin3 (R&D Systems #AF1197, 1:500; Invitrogen #A21447, AlexaFluor647), Iba1 (Wako Laboratory Chemicals #019-19741, 1:500; Invitrogen #A21206, AlexaFluor488), GFAP-Cy3 (Millipore Sigma #C9205, 1:500), GluN2A (Invitrogen #480031; Invitrogen #A11008 AlexaFluor488).

Techniques: Expressing, Marker, Labeling, Battery