Journal: Nature Communications

Article Title: Priming versus propagating: distinct immune effects of alpha- versus beta-particle emitting radiopharmaceuticals when combined with immune checkpoint inhibition in mice

doi: 10.1038/s41467-026-68834-1

Figure Lengend Snippet: A Confirmation of CD8 + depletion by flow cytometry. Gating strategy: Supplementary Fig. . B – K MC38 tumor-bearing mice received 2 Gy 90 Y-, 177 Lu-, or 225 Ac-NM600 + ICI (days −3/0/3) + αCD8 (anti-CD8 on days −5/0/5), RPT + ICI, RPT alone, or αCD8 alone. Effects of CD8 + depletion on tumor growth ( B – H ) and overall survival ( I – K ) are shown. L Flow cytometry treatment scheme. MC38 tumor-bearing mice were randomized to receive either 2 Gy 90 Y-, 177 Lu-, or 225 Ac-NM600 + ICI (days −3/0/3), ICI alone, or untreated control (No Tx). M , N Tumors were harvested on days −3, 4, 8, or 11 and dissociated. The tumor immune infiltrate was analyzed by flow cytometry. Gating strategy: Supplementary Fig. . O Co-culture experiment scheme. Created https://BioRender.com/22ha1p4 . P , Q TNF and IFNγ expression were measured by intracellular flow cytometry staining and analysis of peripheral CD8 + effector memory cells (CD8 + CD44 + CD62L − ) harvested from naïve C57BL/6 mice or treated MC38 tumor-bearing mice following 24 h of co-culture with MC38 or no tumor cells. Gating strategy: Supplementary Fig. . A – H ; M , N ; P , Q Results of one experiment. I – K Results of two independent experiments. A N = 15: depleted; n = 7: IgG2b control. B – H N = 7: 90 Y-, 177 Lu-, 225 Ac-NM600 ± ICI; n = 5: 90 Y-, 177 Lu-, 225 Ac-NM600 + ICI + αCD8, αCD8. I – K N = 17: 90 Y-, 177 Lu-, 225 Ac-NM600 ± ICI; n = 10: αCD8; n = 5: 90 Y-, 177 Lu-, 225 Ac-NM600 + ICI + αCD8. M , N N = 5/treatment group and timepoint except n = 4: 90 Y-NM600 + ICI Day 4; 90 Y-, 177 Lu-, 225 Ac-NM600 + ICI Day 11. P , Q N = 5: naïve, 2 Gy 90 Y-NM600 + ICI; n = 6: ICI, 2 Gy 177 Lu-NM600 + ICI; n = 7: 2 Gy 225 Ac-NM600 + ICI. Unpaired two-tailed t-test was used to compare CD8 + frequency between groups. One-way ANOVA with Tukey’s HSD post hoc test was used to compare flow cytometry results between treatment groups for each marker and timepoint and TNF or IFNγ expression between treatment groups within each co-culture condition. Linear mixed models were used to compare tumor volumes over time between various treatment groups. Statistical testing of the pairwise contrasts was adjusted for multiple comparisons using Tukey’s method. Log-rank test was used to compare survival. Error bars are SEM. O Created in BioRender. Jin, W. (2026) https://BioRender.com/22ha1p4 .

Article Snippet: CD8 + T cell depletion was performed by injection of anti-murine CD8α (Rat IgG2b, κ; Clone 2.43; BioXCell, Cat # BE0061) or IgG2b control (Rat IgG2b, κ; Clone LTF-2; BioXCell, Cat # BE0090) by 300 μg intraperitoneal injection on days −5, 0, and 5.

Techniques: Flow Cytometry, Control, Co-Culture Assay, Expressing, Staining, Two Tailed Test, Marker

Journal: Nature Communications

Article Title: Priming versus propagating: distinct immune effects of alpha- versus beta-particle emitting radiopharmaceuticals when combined with immune checkpoint inhibition in mice

doi: 10.1038/s41467-026-68834-1

Figure Lengend Snippet: MC38 tumor-bearing mice received IgG2b control on days −5/0/5 (IgG2b), or 0, 0.2 Gy (0.4625 kBq), 2 Gy (4.625 kBq), 8 Gy (18.5 kBq) 225 Ac-NM600 on day 1 ± ICI on days −3/0/3. A Body weights. B Complete blood counts on day 23. WBC white blood cells, LYM lymphocytes, PLT platelets, HGB hemoglobin, RBC red blood cells. C TNF and IFNγ on peripheral CD8 + effector memory cells (CD8 + CD44 + CD62L − ) from naïve or MC38 tumor-bearing mice receiving the indicated treatment and following 24 h of co-culture with MC38 or no tumor cells (from Fig. ). D – L MC38 tumor-bearing mice received 8 Gy 225 Ac-NM600 + ICI, 8 Gy 225 Ac-NM600, ICI, or were untreated (No Tx). D Experimental scheme. Created with BioRender.com. Tumor growth ( E ) and overall survival ( F ) are shown. G Complete response (CR) rates ( H ) and percent of CR mice and naïve controls rejecting rechallenge with MC38 cells are shown. I – K Flow cytometry of blood from CR (MC38, Ac + ICI) or naïve mice (MC38, naïve). L Flow cytometry of splenocytes from CR 225 Ac-NM600 + ICI mice that did not (Ac + ICI tumor) or did reject rechallenge (Ac + ICI no tumor), CR ICI mice (ICI), naïve C57BL/6 mice following IV injection of MC38 cells (MC38 IV), and completely naïve C57BL/6 mice (Naïve). Gating strategies: Supplementary Figs. – . A , B ; C ; E – L Results of one experiment. A N = 7: all groups. B N = 5: naïve, 2 Gy 90 Y-NM600 + ICI; n = 6: ICI, 2 Gy 177 Lu-NM600 + ICI; n = 7: 0.2, 2, 8 Gy 225 Ac-NM600 + ICI. C N = 5: naïve; n = 6: ICI; n = 7: 0.2, 2, 8 Gy 225 Ac-NM600 + ICI. E – G N = 5/treatment group. H N = 5: 225 Ac-NM600 + ICI, naïve; n = 3: ICI. I – K N = 4/treatment group. L N = 4: MC38 IV, Naïve; n = 3: ICI; n = 2: Ac + ICI tumor, Ac + ICI no tumor. One-way ANOVA with Tukey’s HSD post hoc test was used to compare TNF or IFNγ expression, blood counts, CR rates, rechallenge rejection rates, and spleen flow cytometry. Unpaired two-tailed t-test was used to compare blood flow cytometry. Tumor volumes were compared using linear mixed models adjusted for multiple comparisons using Tukey’s method. Log-rank test was used to compare survival. Error bars are SEM. D Created in BioRender. Berg, T. (2026) https://BioRender.com/llbyoho, .

Article Snippet: CD8 + T cell depletion was performed by injection of anti-murine CD8α (Rat IgG2b, κ; Clone 2.43; BioXCell, Cat # BE0061) or IgG2b control (Rat IgG2b, κ; Clone LTF-2; BioXCell, Cat # BE0090) by 300 μg intraperitoneal injection on days −5, 0, and 5.

Techniques: Control, Co-Culture Assay, Flow Cytometry, IV Injection, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Priming versus propagating: distinct immune effects of alpha- versus beta-particle emitting radiopharmaceuticals when combined with immune checkpoint inhibition in mice

doi: 10.1038/s41467-026-68834-1

Figure Lengend Snippet: Paired TCR sequencing analysis performed on samples from B78 tumor-bearing mice used for single cell RNA sequencing. A – H Data presented are from samples from experiment depicted in Fig. . A UMAP plots of T cells stratified by sample. B TCR clonotype expansion ranges. C UMAP plots of T cells stratified by T cell subtype. D Clonal expansion by treatment group and T cell subtype. E D50 score by treatment group and T cell subtype. F Relative abundance of clonal indices for each treatment group and T cell subtype. G UpSet plot showing shared clonotypes between T cell subtypes for the 225 Ac-NM600 + ICI sample. Red box demarcates CD8 + effector and memory subsets; red arrow demarcates # of shared clonotypes between CD8 + effector and memory subsets (=11). H UpSet plots showing shared clonotypes between T cell subtypes for the 90 Y-, 177 Lu-, EBRT + ICI, and ICI alone samples. For each plot, red arrow demarcates # of shared clonotypes between CD8+ effector and memory subsets (=7 for each sample). A – H Results of one experiment. 3 mice/treatment group pooled into one sample/treatment group for TCR sequencing analysis. Numbers of cells analyzed provided in Supplementary Table . Number of cells analyzed: 90 Y-NM600 + ICI, n = 408; 177 Lu-NM600 + ICI, n = 993; 225 Ac-NM600 + ICI, n = 854; EBRT + ICI, n = 285; ICI, n = 977.

Article Snippet: CD8 + T cell depletion was performed by injection of anti-murine CD8α (Rat IgG2b, κ; Clone 2.43; BioXCell, Cat # BE0061) or IgG2b control (Rat IgG2b, κ; Clone LTF-2; BioXCell, Cat # BE0090) by 300 μg intraperitoneal injection on days −5, 0, and 5.

Techniques: Sequencing, Single Cell, RNA Sequencing

Journal: Nature Communications

Article Title: Priming versus propagating: distinct immune effects of alpha- versus beta-particle emitting radiopharmaceuticals when combined with immune checkpoint inhibition in mice

doi: 10.1038/s41467-026-68834-1

Figure Lengend Snippet: A – D Data presented are from samples from experiment depicted in Fig. . A Pathway enrichment results for CD8 + effector T cells. The bluer the pathway, the lower the adjusted P, and thus the more significantly the pathway is altered. Differential gene expression heat map for interferon γ ( B ) and interferon α ( C ) response genes for CD8 + effector T cells. The color of each box represents the log 2 (fold change) of gene expression of 225 Ac-, 177 Lu-, 90 Y-NM600, or EBRT + ICI compared to ICI alone. D Incidence matrix of differential gene expression between treatment groups of the following: comparing cells where the shared clonotypes between effector and memory CD8 + T cells are between 7 and 11 (as shown in Fig. ), and they are amplified. Red arrow marks interferon-inducible gene, Ifi27l2a , which is upregulated in these shared clonotype CD8 + cell populations treated with 225 Ac-NM600 + ICI over each treatment group. A – D Results of one experiment. 3 mice/treatment group pooled into one sample/treatment group for single cell RNA sequencing TCR sequencing analysis. Benjamini–Hochberg method was used to correct for multiple comparison. Numbers of cells analyzed provided in Supplementary Table A – C 90 Y-NM600 + ICI, n = 6067; 177 Lu-NM600 + ICI, n = 9808; 225 Ac-NM600 + ICI, n = 6503; EBRT + ICI, n = 7045; ICI, n = 11,249 and Supplementary Table D 90 Y-NM600 + ICI, n = 123; 177 Lu-NM600 + ICI, n = 151; 225 Ac-NM600 + ICI, n = 45; EBRT + ICI, n = 36; ICI, n = 100.

Article Snippet: CD8 + T cell depletion was performed by injection of anti-murine CD8α (Rat IgG2b, κ; Clone 2.43; BioXCell, Cat # BE0061) or IgG2b control (Rat IgG2b, κ; Clone LTF-2; BioXCell, Cat # BE0090) by 300 μg intraperitoneal injection on days −5, 0, and 5.

Techniques: Gene Expression, Amplification, Single Cell, RNA Sequencing, Sequencing, Comparison

Journal: Cell metabolism

Article Title: Oligodendrocytes Provide Antioxidant Defense Function for Neurons by Secreting Ferritin Heavy Chain

doi: 10.1016/j.cmet.2020.05.019

Figure Lengend Snippet: Key Resources Table

Article Snippet: rat anti-CD68 (FA-11) , Biorad , Cat# MCA1957T; RRID: AB_2074849.

Techniques: Recombinant, Amplification, Library Quantification, Sequencing, Proliferation Assay, Enzyme-linked Immunosorbent Assay, Mutagenesis, Quantitative RT-PCR, Expressing, Plasmid Preparation, Software

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: αβ T-cell receptors from multiple sclerosis brain lesions show MAIT cell–related features

doi: 10.1212/NXI.0000000000000107

Figure Lengend Snippet: (A) Cluster of expanded Vβ1 + T cells within multiple sclerosis (MS) lesions. Immunohistochemistry for the expanded and persisting Vβ1 clone (green) in clusters of CD8 + T cells (red) in parenchymal MS lesions. Several such clusters were observed. Only very few scattered CD8 − Vβ1 + T cells could be identified in the brain lesion. Nuclei are visualized with 4',6-diamidino-2-phenylindole (white). Scale bar 20 µm. (B) Sequences of paired T-cell receptor (TCR) α and β chains. Single sorted or laser microdissected Vβ1 + CD8 + T cells from peripheral blood or brain sections were submitted to single-cell TCR PCR to identify Vβ1 chains and all possible matching α chains. The V, n(D)n, and J regions are indicated. Amino acids encoded by n(D)n nucleotides are printed in red. The expanded Vβ1-Jβ2.3 β chain (upper line) was found to pair with 4 different α chains. Three α chains were identified from brain lesions, and 1 α chain was found in blood. The α chains expressing the Jα33 (second line) and Jα16 (fifth line) elements were identified in 7 and 6 independent cells, respectively. All α chains share the Vα7.2 element, and even though they do not share the same Jα element, they all show homologous complementarity determining region 3α regions with a conserved valine (V) followed by a positively charged arginine (R) (with only one clone showing a glutamine [Q]), a negatively charged amino acid (D/E), and a relatively large hydrophilic amino acid. One of the α chains (highlighted in red) is the mucosal-associated invariant T (MAIT) cell canonical TCR Vα7.2-CAXXDSNYQLIW-Jα33 chain with 2 N nucleotide–encoded amino acids between Vα7.2 and Jα33 (here VR). The other clones with Jα16, Jα24.1, and Jα58 chains are atypical for MAIT cells, which usually carry Jα33, Jα20, or Jα12. PBMC = peripheral blood mononuclear cell.

Article Snippet: To characterize T-cell infiltrates in sections of MS brain, the following antibodies against cell surface molecules were used: mouse anti-human CD161 (1:5, 191B8, Miltenyi Biotec, Bergisch Gladbach, Germany), mouse anti-human Vα7.2 (1:5, 3C10, BioLegend, San Diego, CA), mouse anti-human CD8α (1:50, LT8, AbD Serotec, Kidlington, UK; labeled with the Cy3 MAb labeling kit, GE Healthcare, Freiburg, Germany), rabbit anti-human CCR7 (1:800, Y59, Abcam, Cambridge, UK), mouse anti-human CD45RA (1:250, HI100, BioLegend), mouse anti-human CD45RO (1:250, UCHL1, BioLegend), fluorescein isothiocyanate (FITC)-labeled mouse anti-human Vβ1 (1:100, BL37.2, Beckman Coulter, Brea, CA), rabbit anti-human CD3 (1:500, Dako, Glostrup, Denmark), and Alexa Fluor 488–conjugated mouse anti-human CD4 (1:50, RPA-T4, eBioscience, San Diego, CA).

Techniques: Immunohistochemistry, Expressing, Clone Assay

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: αβ T-cell receptors from multiple sclerosis brain lesions show MAIT cell–related features

doi: 10.1212/NXI.0000000000000107

Figure Lengend Snippet: Immunofluorescence staining of brain-infiltrating immune cells. All nuclei are stained with 4′,6-diamidino-2-phenylindole (white). Green and red dyes were used. Double-positive cells are therefore shown in yellow. Scale bars 50 µm. (A) Double staining for CD3 (green) and CD8α (red). Most CD3 + T cells coexpress CD8α. (B) CD8 + (red) T cells outnumber CD4 + (green) T cells. (C) Low numbers of CD45RA + (green) CD8 + (red) double-positive T cells in multiple sclerosis (MS) brain tissue. (D) Many CD8 + T cells (red) coexpress CD45RO (green). (E) Naive T cells double-positive for CCR7 (red) and CD45RA (green) are mostly found within blood vessels and are barely detectable in the parenchyma of MS CNS. (F) Effector memory (CD45RO + CCR7 − ) (green arrow) and central memory (CD45RO + CCR + ) (yellow arrow) T cells in MS lesions.

Article Snippet: To characterize T-cell infiltrates in sections of MS brain, the following antibodies against cell surface molecules were used: mouse anti-human CD161 (1:5, 191B8, Miltenyi Biotec, Bergisch Gladbach, Germany), mouse anti-human Vα7.2 (1:5, 3C10, BioLegend, San Diego, CA), mouse anti-human CD8α (1:50, LT8, AbD Serotec, Kidlington, UK; labeled with the Cy3 MAb labeling kit, GE Healthcare, Freiburg, Germany), rabbit anti-human CCR7 (1:800, Y59, Abcam, Cambridge, UK), mouse anti-human CD45RA (1:250, HI100, BioLegend), mouse anti-human CD45RO (1:250, UCHL1, BioLegend), fluorescein isothiocyanate (FITC)-labeled mouse anti-human Vβ1 (1:100, BL37.2, Beckman Coulter, Brea, CA), rabbit anti-human CD3 (1:500, Dako, Glostrup, Denmark), and Alexa Fluor 488–conjugated mouse anti-human CD4 (1:50, RPA-T4, eBioscience, San Diego, CA).

Techniques: Immunofluorescence, Staining, Double Staining

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: αβ T-cell receptors from multiple sclerosis brain lesions show MAIT cell–related features

doi: 10.1212/NXI.0000000000000107

Figure Lengend Snippet: Double fluorescence immunohistochemistry identifies brain-infiltrating mucosal-associated invariant T (MAIT) cells in multiple sclerosis (MS) lesions of patient A. Nuclei are stained with 4',6-diamidino-2-phenylindole (white). Green and red dyes were used. Double-positive cells are therefore shown in yellow. Scale bars 20 µm. (A) T cells expressing the T-cell receptor Vα7.2 (red) and Vβ1 chains (green). This combination (see ) was identified by single-cell PCR. (B) Most Vα7.2 + (green) T cells belong to the CD8 + (red) T-cell subset. (C) MAIT cells expressing Vα7.2 (red) and CD161 (green) in the parenchyma of MS brain. (D) The vast majority of CD161 + (green) cells in MS CNS coexpress CD8α (red).

Article Snippet: To characterize T-cell infiltrates in sections of MS brain, the following antibodies against cell surface molecules were used: mouse anti-human CD161 (1:5, 191B8, Miltenyi Biotec, Bergisch Gladbach, Germany), mouse anti-human Vα7.2 (1:5, 3C10, BioLegend, San Diego, CA), mouse anti-human CD8α (1:50, LT8, AbD Serotec, Kidlington, UK; labeled with the Cy3 MAb labeling kit, GE Healthcare, Freiburg, Germany), rabbit anti-human CCR7 (1:800, Y59, Abcam, Cambridge, UK), mouse anti-human CD45RA (1:250, HI100, BioLegend), mouse anti-human CD45RO (1:250, UCHL1, BioLegend), fluorescein isothiocyanate (FITC)-labeled mouse anti-human Vβ1 (1:100, BL37.2, Beckman Coulter, Brea, CA), rabbit anti-human CD3 (1:500, Dako, Glostrup, Denmark), and Alexa Fluor 488–conjugated mouse anti-human CD4 (1:50, RPA-T4, eBioscience, San Diego, CA).

Techniques: Fluorescence, Immunohistochemistry, Staining, Expressing

Journal: Neurology® Neuroimmunology & Neuroinflammation

Article Title: αβ T-cell receptors from multiple sclerosis brain lesions show MAIT cell–related features

doi: 10.1212/NXI.0000000000000107

Figure Lengend Snippet: Analysis of the T-cell receptor (TCR) Vα7.2 repertoire of patient A by pyrosequencing shows oligoclonal T-cell expansions in different samples. Each pie chart represents the total number of nucleotide sequences found in the Vα7.2 repertoire of a certain compartment and each sector represents one distinct T-cell clone. We compared (A) samples from CNS tissue from the biopsy taken in 1996, (B) Vα7.2 + CD161 + mucosal-associated invariant T (MAIT) cells, (C) Vα7.2 + CD161 − cells, (D) CD8 + cells, and (E) CD4 + T cells from peripheral blood taken in 2013 and 2014. For each population we list the designation of the Jα elements and their relative percentage. Clones carrying the MAIT canonical TCR α chain (CAXXDSNYQLIW) are marked in bright blue, and clones containing the noncanonical α chains characterized by Jα12 (CAXXDSSYKLIF) and Jα20 (CAVXXDYKLSF) are marked in dark blue and light blue, respectively. (F) Percentages of identical complementarity determining region 3α amino acid sequences within the TCR Vα7.2 + repertoire between the different samples defined above (A–E). The numbers are percentages indicating how often a particular sequence detected in one sample was also found in another sample. The greatest overlap was between Vα7.2 + CD161 + and CD8 + T cells from peripheral blood, but there was also significant overlap between the CNS sample and the Vα7.2 + CD161 + sample from 2013 to 2014, as highlighted by the red and yellow colors. PBMC = peripheral blood mononuclear cell.

Article Snippet: To characterize T-cell infiltrates in sections of MS brain, the following antibodies against cell surface molecules were used: mouse anti-human CD161 (1:5, 191B8, Miltenyi Biotec, Bergisch Gladbach, Germany), mouse anti-human Vα7.2 (1:5, 3C10, BioLegend, San Diego, CA), mouse anti-human CD8α (1:50, LT8, AbD Serotec, Kidlington, UK; labeled with the Cy3 MAb labeling kit, GE Healthcare, Freiburg, Germany), rabbit anti-human CCR7 (1:800, Y59, Abcam, Cambridge, UK), mouse anti-human CD45RA (1:250, HI100, BioLegend), mouse anti-human CD45RO (1:250, UCHL1, BioLegend), fluorescein isothiocyanate (FITC)-labeled mouse anti-human Vβ1 (1:100, BL37.2, Beckman Coulter, Brea, CA), rabbit anti-human CD3 (1:500, Dako, Glostrup, Denmark), and Alexa Fluor 488–conjugated mouse anti-human CD4 (1:50, RPA-T4, eBioscience, San Diego, CA).

Techniques: Clone Assay, Sequencing

Journal: Cell

Article Title: Human neutralizing antibodies against SARS-CoV-2 require intact Fc effector functions for optimal therapeutic protection

doi: 10.1016/j.cell.2021.02.026

Figure Lengend Snippet: Confirmation of cellular depletions, related to Figure 6 (A) Representative flow cytometry plots of monocytes and neutrophils from peripheral blood at 8 dpi following intraperitoneal injection of a depleting anti-CCR2 mAb or isotype control mAb and frequency of Ly6C hi monocytes and neutrophils in blood at 8 dpi following anti-CCR2 or isotype control mAb administration in isotype control or COV2-2050-treated mice (two experiments, n = 6 per group). (B) Representative flow cytometry plots of peripheral blood at 8 dpi following intraperitoneal injection of a depleting anti-Ly6G mAb or isotype control mAb and frequency of Ly6C hi monocytes and mature Ly6G + neutrophils in blood at 8 dpi following anti-Ly6G or isotype control mAb administration in isotype control or COV2-2050-treated mice (two experiments, n = 5-6 per group). In the groups treated with anti-Ly6G, neutrophils were identified as CD11b + Ly6B + Ly6C int cells. Red dots (neutrophils) and monocytes (blue dots) correspond to the same population identified in both plots (neutrophils: CD11b + Ly6C + Ly6G + Ly6C int and monocytes: CD11b + Ly6C + Ly6G - Ly6C high ). (C) Representative flow cytometry plots of peripheral blood at 8 dpi following intraperitoneal injection of a depleting anti-NK1.1 mAb or isotype control mAb. Also shown is the frequency of NK cells in blood at 8 dpi following anti-NK1.1 or isotype control mAb administration in isotype control or COV2-2050-treated mice (two experiments, n = 4-5 per group). (D) Representative flow cytometry plots of splenocytes gated on CD4 + and CD8 + T cells at 8 dpi following intraperitoneal injection of a depleting anti-CD8α mAb or isotype control mAb (left). Frequency of CD8 + T cells in the spleen at 8 dpi following anti-CD8α or isotype control mAb administration in isotype mAb control or COV2-2050-treated mice (right) (two experiments, n = 4-5 per group). (E) Representative flow cytometry plots of splenocytes gated to cDC1s at 8 dpi following intraperitoneal injection of a depleting anti-CD8α mAb or isotype control mAb (left). Frequency of cDC1 cells in the spleen at 8 dpi following anti-CD8α or isotype control mAb administration in isotype control mAb or COV2-2050-treated mice (right) (two experiments, n = 4-5 per group).

Article Snippet: InVivoPlus anti-mouse CD8α , BioXCell , RRID: AB_10950145.

Techniques: Flow Cytometry, Injection

Journal: Cell

Article Title: Human neutralizing antibodies against SARS-CoV-2 require intact Fc effector functions for optimal therapeutic protection

doi: 10.1016/j.cell.2021.02.026

Figure Lengend Snippet:

Article Snippet: InVivoPlus anti-mouse CD8α , BioXCell , RRID: AB_10950145.

Techniques: Recombinant, Expressing, Isolation, Staining, Amplification, Software, Plasmid Preparation

Journal: Frontiers in Veterinary Science

Article Title: The immunomodulation–immunogenicity balance of equine Mesenchymal Stem Cells (MSCs) is differentially affected by the immune cell response depending on inflammatory licensing and major histocompatibility complex (MHC) compatibility

doi: 10.3389/fvets.2022.957153

Figure Lengend Snippet: Primers used for gene expression analysis by RT-qPCR.

Article Snippet: For IL6 analysis (Equine IL-6 DuoSet ELISA, R&D Systems, REF: DY1886), baseline and control supernatants were diluted 1:1 in the reagent diluent, and supernatants from co-cultures were not diluted.

Techniques: Gene Expression, Sequencing, Amplification

Journal: Frontiers in Veterinary Science

Article Title: The immunomodulation–immunogenicity balance of equine Mesenchymal Stem Cells (MSCs) is differentially affected by the immune cell response depending on inflammatory licensing and major histocompatibility complex (MHC) compatibility

doi: 10.3389/fvets.2022.957153

Figure Lengend Snippet: Interleukin 6 (IL6) gene expression and secretion by equine mesenchymal stem cells (MSCs) in the different scenarios. (A) IL6 mRNA relative expression and (C) IL6 secretion (pg/mL) before (baseline; orange bars) and after equine MSC-naive (light blue bars) and MSC-primed (dark blue bars) were exposed in vitro to phytohemagglutinin (PHA)-activated peripheral blood lymphocytes (PBLs) (immunosuppression assays). (B) IL6 mRNA relative expression and (D) IL6 secretion before (baseline; orange bars) and after MSC-naive (light green bars) and MSC-primed (dark green bars) were exposed in vitro to resting PBLs [modified one-way mixed lymphocyte reaction (MLR) assays]. Co-cultures of MSCs and PBLs were autologous ( n = 3), allogeneic, matched ( n = 8), or mismatched ( n = 7) for the major histocompatibility complex. Changes in gene expression are represented as mean ± S.E.M of the relative mRNA expression, using baseline MSC-naive as reference sample (light orange bar, value 1). Concentration of IL6 in the supernatant from the different conditions is represented as mean ± S.E.M (pg/mL). Significant differences of each condition compared with the baseline MSC-naive (light orange bar) are represented by hashes (#) above the corresponding bar ( # p < 0.05; #### p < 0.0001). Significant differences compared with the baseline MSC-primed (dark orange bar) are represented by a cross (+) above the corresponding bar ( + p < 0.05; ++ p < 0.01). Significant differences between experimental conditions are represented by a squared line with an asterisk ( * p < 0.05; *** p < 0.001).

Article Snippet: For IL6 analysis (Equine IL-6 DuoSet ELISA, R&D Systems, REF: DY1886), baseline and control supernatants were diluted 1:1 in the reagent diluent, and supernatants from co-cultures were not diluted.

Techniques: Gene Expression, Expressing, In Vitro, Modification, Immunopeptidomics, Concentration Assay

Journal: The Journal of Biological Chemistry

Article Title: Double Deficiency of Tetraspanins CD9 and CD81 Alters Cell Motility and Protease Production of Macrophages and Causes Chronic Obstructive Pulmonary Disease-like Phenotype in Mice * S⃞

doi: 10.1074/jbc.M801902200

Figure Lengend Snippet: TSA or CSE down-regulates CD9 and CD81 while up-regulating MMPs in RAW264.7 macrophages. A, RAW264.7 cells were cultured in the absence or presence of TSA, theophylline, and dexamethasone for 48 h. Expressions of CD9, CD81, and integrin β1 were examined by immunoblotting using whole cell lysates. Anti-actin blots confirm equal amounts of protein loaded in each lane. B, RAW264.7 was cultured in the absence (filled histograms) or presence (open histograms) of TSA. Surface expressions of CD9, CD81, and integrin β1 were analyzed by flow cytometry. C, A549 alveolar epithelial cells were cultured in the absence or presence of TSA. CD9 and CD81 were immunoblotted. D, RAW264.7 was cultured in the absence or presence of 0.1% CSE for 48 h. CD9, CD81, and integrin β1 were immunoblotted. E, RAW264.7 was cultured in the absence or presence of TSA. Expressions of MMP-2, MMP-9, and MMP-12 were analyzed by RT-PCR. β-Actin amplification was internal control.

Article Snippet: Cell lysates were separated by SDS-PAGE, transferred to polyvinylidene difluoride membranes, and probed with rat anti-mouse CD9 (KMC8) and integrin β1 (KMI6) mAbs (BD Biosciences) and hamster anti-mouse CD81 mAb (Eat2; AbD Serotec, Oxford, UK).

Techniques: Cell Culture, Western Blot, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Amplification, Control

Journal: The Journal of Biological Chemistry

Article Title: Double Deficiency of Tetraspanins CD9 and CD81 Alters Cell Motility and Protease Production of Macrophages and Causes Chronic Obstructive Pulmonary Disease-like Phenotype in Mice * S⃞

doi: 10.1074/jbc.M801902200

Figure Lengend Snippet: mAbs or siRNA to CD9 and CD81 enhance MMP production and suppress motility of RAW264.7 cells. A, RAW264.7 cells were cultured for 24 h in the absence (-) or presence of KMC8 plus 2F7 (+). mRNA was extracted, and RT-PCR was performed for expressions of MMPs and TIMPs. B, RAW264.7 cells were cultured for 24 h in the absence (-) or presence of the indicated mAbs. MMP-9 activity in culture supernatant was examined by gelatin zymography (lower) and quantified by densitometry (upper). The gelatin zymography is from one representative of three similar experiments. C, RAW264.7 was transfected with siRNAs against CD9 or CD81. Decrease in CD9 or CD81 was shown in immunoblotting (upper). MMP-9 activity in supernatants of 24-hour culture was examined by gelatin zymography (lower). D, RAW264.7 cells were applied into the upper chamber of FN-precoated Transwells in the absence (-) or presence of the indicated mAbs. DMEM containing 10% FBS was added to the lower chamber. After 4 h, cells migrating to the lower surface of the membrane were counted after Diff-Quick stain. Bars represent the mean ± S.E. *, p < 0.05 versus IgG; **, p < 0.05 versus KMC8. KMC8, anti-CD9; 2F7, anti-CD81; HMβ1-1, anti-integrin β1.

Article Snippet: Cell lysates were separated by SDS-PAGE, transferred to polyvinylidene difluoride membranes, and probed with rat anti-mouse CD9 (KMC8) and integrin β1 (KMI6) mAbs (BD Biosciences) and hamster anti-mouse CD81 mAb (Eat2; AbD Serotec, Oxford, UK).

Techniques: Cell Culture, Reverse Transcription Polymerase Chain Reaction, Activity Assay, Zymography, Transfection, Western Blot, Membrane, Diff-Quik, Staining

Journal: The Journal of Biological Chemistry

Article Title: Double Deficiency of Tetraspanins CD9 and CD81 Alters Cell Motility and Protease Production of Macrophages and Causes Chronic Obstructive Pulmonary Disease-like Phenotype in Mice * S⃞

doi: 10.1074/jbc.M801902200

Figure Lengend Snippet: CD9/CD81 DKO mice develop pulmonary emphysema. A, histological lung sections from DKO mice and WT littermates at 3 and 10 weeks of age were stained with hematoxylin and eosin. Bar, 50 μm. B, chord length measured from the WT and DKO lungs. C and D, functional tests of the lung. Lung compliance (C) and functional residual capacity (FRC) (D) of mice at 23 weeks of age were measured with a whole body plethysmograph. Values were normalized to body weight. At least three mice were used for each group. Bars represent the mean ± S.E. *, p < 0.05 versus WT.

Article Snippet: Cell lysates were separated by SDS-PAGE, transferred to polyvinylidene difluoride membranes, and probed with rat anti-mouse CD9 (KMC8) and integrin β1 (KMI6) mAbs (BD Biosciences) and hamster anti-mouse CD81 mAb (Eat2; AbD Serotec, Oxford, UK).

Techniques: Staining, Functional Assay

Journal: The Journal of Biological Chemistry

Article Title: Double Deficiency of Tetraspanins CD9 and CD81 Alters Cell Motility and Protease Production of Macrophages and Causes Chronic Obstructive Pulmonary Disease-like Phenotype in Mice * S⃞

doi: 10.1074/jbc.M801902200

Figure Lengend Snippet: CD9/CD81 DKO mice display body weight loss, kyphosis, and osteopenia. A, whole body radiographs of WT and DKO mice at 3 and 30 weeks of age. Bars, 10 mm. B, time course of body weight of WT and DKO littermates. C, toluidine blue stain of longitudinal sections of proximal tibia at 8 weeks of age. Note that cortical bone of the DKO mouse is thinner than that of the WT littermate (arrowheads). Bar, 250 μm. D, pQCT images of femoral diaphysis at 8 weeks of age (upper). Mineral densities are shown as different colors according to the standard mineral density gradients. Note the reduction in cortical thickness of the DKO mouse (arrowheads). Total mineral content and strength strain index (SSI) were also determined (lower). Trabecular parameters were not different between WT and DKO mice (data not shown). Values represent the mean ± S.E. *, p < 0.05 versus WT.

Article Snippet: Cell lysates were separated by SDS-PAGE, transferred to polyvinylidene difluoride membranes, and probed with rat anti-mouse CD9 (KMC8) and integrin β1 (KMI6) mAbs (BD Biosciences) and hamster anti-mouse CD81 mAb (Eat2; AbD Serotec, Oxford, UK).

Techniques: Staining

Journal: iScience

Article Title: Kaempferol attenuates hyperuricemia combined with gouty arthritis via urate transporters and NLRP3/NF-κB pathway modulation

doi: 10.1016/j.isci.2024.111186

Figure Lengend Snippet:

Article Snippet: Mouse IL-6 ELISA Kit , Boster Bio Technology , Cat# EK0411.

Techniques: Recombinant, Lysis, Protease Inhibitor, ROS Assay, LDH Cytotoxicity Assay, Multiple Displacement Amplification, GSH Assay, Enzyme-linked Immunosorbent Assay, Software