Journal: Cancer immunology research

Article Title: Aggressive Mammary Cancers Lacking Lymphocytic Infiltration Arise in Irradiated Mice and Can be Prevented by Dietary Intervention

doi: 10.1158/2326-6066.CIR-19-0253

Figure Lengend Snippet: Antibodies for Opal Staining

Article Snippet: Set 4 , CD33 (NBP2–37388, Novus) , Opal Polymer HRP Ms+Rb (ARH1001EA, PerkinElmer) , Opal 520 (NEL811001KT, PerkinElmer).

Techniques: Polymer

Journal: Glia

Article Title: Deletion of Alzheimer's disease-associated CD33 results in an inflammatory human microglia phenotype.

doi: 10.1002/glia.23968

Figure Lengend Snippet: FIGURE 1 CD33 knockout (CD33−/−) and lentiviral shRNA-mediated PTPN6 knockdown in THP1 macrophages. (a) Exemplary flow cytometry histogram graph showing expression levels of CD33 in wild type (WT) and CD33−/−macrophages. CD33 expression is absent in CD33−/−

Article Snippet: The differentiated THP1 macrophages were treated with either 10 μg/ml anti-FcγRI (CD64) antibody (clone 10.1, Santa Cruz Biotechnology #SC-1184), 10 μg/ml anti-FcγRI antibody combined with 10 μg/ml mouse IgG1 isotype control or combined with 10 μg/ml mouse anti-CD33 antibody (clone 1C7/1, Cedarlane #CL7627AP) for 5 min (SYK detection) or 10 min (ERK1/2 detection) at 37 C. CD33 antibody clone 1C7/1 was previously described as putative agonistic antibody (Taylor et al., 1999).

Techniques: Knock-Out, shRNA, Knockdown, Flow Cytometry, Expressing

Journal: Glia

Article Title: Deletion of Alzheimer's disease-associated CD33 results in an inflammatory human microglia phenotype.

doi: 10.1002/glia.23968

Figure Lengend Snippet: FIGURE 2 Transcriptome analysis of CD33 knockout and PTPN6 knockdown in THP1 macrophages. (a) Principal component analysis (PCA) of THP1 macrophage RNA sequencing. PCA resulted in four clusters representing the four individual groups wild type (WT) shCTRL, WT shPTPN6, CD33−/−shCTRL CD33−/−shPTPN6. (b) Heatmap of the top 200 most varying genes with hierarchical clustering for rows and columns stressed that the most extreme differences were between WT and CD33−/−. Minor changes were observed for PTPN6 knockdown. (c) Dotblot of KEGG pathway enrichment analysis revealed an upregulation of the inflammation-related transcription profile in comparisons 1–3 (i.e., 1. WT shCTRL vs. CD33−/−shCTRL, 2. WT shPTPN6 vs. CD33−/−shPTPN6, 3. WT shCTRL vs. WT shPTPN6) but not in comparison 4 (CD33−/−shCTRL vs. CD33−/−shPTPN6)

Article Snippet: The differentiated THP1 macrophages were treated with either 10 μg/ml anti-FcγRI (CD64) antibody (clone 10.1, Santa Cruz Biotechnology #SC-1184), 10 μg/ml anti-FcγRI antibody combined with 10 μg/ml mouse IgG1 isotype control or combined with 10 μg/ml mouse anti-CD33 antibody (clone 1C7/1, Cedarlane #CL7627AP) for 5 min (SYK detection) or 10 min (ERK1/2 detection) at 37 C. CD33 antibody clone 1C7/1 was previously described as putative agonistic antibody (Taylor et al., 1999).

Techniques: Knock-Out, Knockdown, RNA Sequencing, Comparison

Journal: Glia

Article Title: Deletion of Alzheimer's disease-associated CD33 results in an inflammatory human microglia phenotype.

doi: 10.1002/glia.23968

Figure Lengend Snippet: FIGURE 3 Activation of SYK and ERK1/2 signaling pathways as a consequence of CD33 knockout in THP1 macrophages and human-induced pluripotent stem cell-derived microglia (iPSdMiG). (a) Western blot showing phosphorylated ERK1/2 (pERK1/2, top) and total ERK1/2 (tERK1/2, bottom) in wild type (WT) and CD33−/−macrophages. One representative image of five independent experiments is shown. (b) Quantification of pERK1/2 to tERK1/2 ratio in WT (black) and CD33−/−THP1 macrophages (dark gray). Knockout of CD33 resulted in an increase in pERK1/2 in THP1 macrophages. FcγRI antibody treatment tended to increase pERK1/2 levels slightly in WT THP1 macrophages. CD33 antibody treatment following FcγRI antibody treatment tended to decrease the FcγRI antibody-driven increase in pERK1/2 to tERK1/2 levels only in WT THP1 macrophages. Data are shown as mean + SEM (n = 3–5). *p ≤.05 determined by two-way analysis of variance (ANOVA) followed by Bonferroni post hoc test. (c) Western blot showing phosphorylated SYK (pSYK, top) and total SYK (tSYK, bottom) in WT and CD33−/−macrophages. One representative image of three independent experiments is shown. (d) Quantification of pSYK to tSYK ratio in WT (black) and CD33−/−THP1 macrophages (dark gray). Knockout of CD33 resulted in an increase in pSYK in THP1 macrophages. FcγRI antibody treatment tended to increase pSYK levels slightly and co-treatment with a CD33 antibody tended to counteract this increase, both only in WT THP1 macrophages. Data are shown as mean + SEM (n = 3). *p ≤.05 determined by two-way ANOVA followed by Bonferroni post hoc test. (e) SYK phosphorylation relative to total SYK assessed by AlphaLISA. Untreated iPSdMiG revealed an increased pSYK/tSYK ratio followed by knockout of CD33 (dark gray) or expression of CD33ΔE2 (light gray) compared to WT iPSdMiG (black). Data are shown as mean + SEM (n = 6). **p ≤.01, *p ≤.05 determined by one-way ANOVA followed by Bonferroni post hoc test. (f) Treatment of iPSdMiG with activating antibodies for TREM2 resulted in an increased pSYK/tSYK ratio in WT (black), CD33−/−(dark gray) and CD33ΔE2 iPSdMiG (light gray) compared to isotype antibody-treated controls. Further, pSYK/tSYK was sharply increased in CD33−/−iPSdMiG following anti-TREM2 treatment. Data are shown as mean + SEM (n = 3). ***p ≤.001, *p ≤.05 determined by two-way ANOVA followed by Bonferroni post hoc test

Article Snippet: The differentiated THP1 macrophages were treated with either 10 μg/ml anti-FcγRI (CD64) antibody (clone 10.1, Santa Cruz Biotechnology #SC-1184), 10 μg/ml anti-FcγRI antibody combined with 10 μg/ml mouse IgG1 isotype control or combined with 10 μg/ml mouse anti-CD33 antibody (clone 1C7/1, Cedarlane #CL7627AP) for 5 min (SYK detection) or 10 min (ERK1/2 detection) at 37 C. CD33 antibody clone 1C7/1 was previously described as putative agonistic antibody (Taylor et al., 1999).

Techniques: Activation Assay, Protein-Protein interactions, Knock-Out, Derivative Assay, Western Blot, Phospho-proteomics, Expressing

Journal: Glia

Article Title: Deletion of Alzheimer's disease-associated CD33 results in an inflammatory human microglia phenotype.

doi: 10.1002/glia.23968

Figure Lengend Snippet: FIGURE 4 Inflammatory cytokine gene transcription is elevated after CD33 knockout or PTPN6 knockdown. (a) Semiquantitative real-time polymerase chain reaction (PCR) of TNFA gene transcription in THP1 macrophages. Gene transcription levels of TNFA were unchanged in untreated wild type (black) and CD33−/−(dark gray) THP1 macrophages regardless of PTPN6 knockdown (shPTPN6). Following LPS treatment TNFA levels were increased. (b) IL1B mRNA levels in THP1 macrophages determined by semiquantitative real-time PCR. Knockout of CD33 increased gene transcription of IL1B in THP1 macrophages (dark gray), which was further elevated by treatment with LPS. IL1B levels tended to be increased after PTPN6 knockdown (shPTPN6) in wild-type THP1 macrophages (black). (c) IL8 gene transcription in THP1 macrophages analyzed via semiquantitative real-time PCR. IL8 mRNA levels were increased after knockout of CD33 (dark gray) and elevated in all cell lines after LPS treatment. PTPN6 knockdown (shPTPN6) attenuated the increase in IL8 transcription in CD33−/−THP1 macrophages. (d) Semiquantitative real-time PCR of IL10 mRNA levels in THP1 macrophages. Gene transcription levels of IL10 were increased in CD33−/−THP1 macrophages (dark gray). Knockdown of PTPN6 decreased IL10 gene transcription below the control vector levels (shCTRL) in both, untreated and LPS-treated THP1 macrophages. Data are shown as mean + SEM (n = 3–7). ***p ≤.001, **p ≤.01, *p ≤.05 determined by two-way analysis of variance (ANOVA) followed by Bonferroni post hoc test. (e–i) Gene transcription levels of the inflammatory cytokines TNFA, IL1B, IL8, IL10, and IL6 analyzed in wild type (WT) (black), CD33−/−(dark gray) and CD33ΔE2 human-induced pluripotent stem cell-derived microglia (iPSdMiG) (light gray) by semiquantitative real-time PCR. Knockout of CD33 as well as expression of CD33ΔE2 in iPSdMiG resulted in constitutively elevated mRNA levels of (e) TNFA, (f) IL1B, (g) IL8, (h) IL10, and (i) IL6. TNFA, IL10 and IL6 mRNA levels were decreased in CD33ΔE2 iPSdMiG compared to CD33−/−

Article Snippet: The differentiated THP1 macrophages were treated with either 10 μg/ml anti-FcγRI (CD64) antibody (clone 10.1, Santa Cruz Biotechnology #SC-1184), 10 μg/ml anti-FcγRI antibody combined with 10 μg/ml mouse IgG1 isotype control or combined with 10 μg/ml mouse anti-CD33 antibody (clone 1C7/1, Cedarlane #CL7627AP) for 5 min (SYK detection) or 10 min (ERK1/2 detection) at 37 C. CD33 antibody clone 1C7/1 was previously described as putative agonistic antibody (Taylor et al., 1999).

Techniques: Knock-Out, Knockdown, Real-time Polymerase Chain Reaction, Control, Plasmid Preparation, Derivative Assay, Expressing

Journal: Glia

Article Title: Deletion of Alzheimer's disease-associated CD33 results in an inflammatory human microglia phenotype.

doi: 10.1002/glia.23968

Figure Lengend Snippet: FIGURE 5 Increased gene transcription of INPP5D (SHIP1) caused by loss of CD33.(a + b) Gene transcription of SIRPA measured by semiquantitative real-time polymerase chain reaction (qRT-PCR). SIRPA mRNA levels were increased in CD33−/−(dark gray) and PTPN6 knockdown (shPTPN6) wild type (WT) THP1 macrophages (black). PTPN6 knockdown in CD33−/−

Article Snippet: The differentiated THP1 macrophages were treated with either 10 μg/ml anti-FcγRI (CD64) antibody (clone 10.1, Santa Cruz Biotechnology #SC-1184), 10 μg/ml anti-FcγRI antibody combined with 10 μg/ml mouse IgG1 isotype control or combined with 10 μg/ml mouse anti-CD33 antibody (clone 1C7/1, Cedarlane #CL7627AP) for 5 min (SYK detection) or 10 min (ERK1/2 detection) at 37 C. CD33 antibody clone 1C7/1 was previously described as putative agonistic antibody (Taylor et al., 1999).

Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Knockdown

Journal: Glia

Article Title: Deletion of Alzheimer's disease-associated CD33 results in an inflammatory human microglia phenotype.

doi: 10.1002/glia.23968

Figure Lengend Snippet: FIGURE 7 Knockout of CD33 increased radical production in human macrophages and microglia. (a) Production of radicals in THP1 macrophages was assessed by dihydroethidium (DHE) staining and confocal microscopy. Radical production was increased by Aβ1-42 in all four cell lines indicated by DHE staining intensity. Thereby, Aβ1-42-induced radical production in CD33−/−shCTRL macrophages (dark gray) was enhanced compared to the corresponding wild-type (black) shCTRL line. Treatment with scavengers SOD1 and Trolox attenuated the Aβ1-42- driven effect to untreated levels. Data are shown as mean + SEM (n = 4). ***p ≤.001, **p ≤.01, *p ≤.05 determined by two-way analysis of variance (ANOVA) followed by Bonferroni post hoc test. (b) Reactive oxygen species (ROS) production in human-induced pluripotent stem cell- derived microglia (iPSdMiG) analyzed by DHE staining. Wild type (WT) (black), CD33−/−(dark gray) and CD33ΔE2 (light gray) iPSdMiG exhibited similar constitutive ROS production. Treatment of the phagocytes with bacterial particles from S. aureus resulted in an elevated production of ROS in all iPSdMiG lines with a higher magnitude in CD33−/−iPSdMiG. Treatment with the scavenger NAC decreased ROS production to untreated levels. Data are shown as mean + SEM (n = 3–6). ***p ≤.001, **p ≤.01 determined by two-way ANOVA followed by Bonferroni post hoc test

Article Snippet: The differentiated THP1 macrophages were treated with either 10 μg/ml anti-FcγRI (CD64) antibody (clone 10.1, Santa Cruz Biotechnology #SC-1184), 10 μg/ml anti-FcγRI antibody combined with 10 μg/ml mouse IgG1 isotype control or combined with 10 μg/ml mouse anti-CD33 antibody (clone 1C7/1, Cedarlane #CL7627AP) for 5 min (SYK detection) or 10 min (ERK1/2 detection) at 37 C. CD33 antibody clone 1C7/1 was previously described as putative agonistic antibody (Taylor et al., 1999).

Techniques: Knock-Out, Staining, Confocal Microscopy, Derivative Assay

Journal: Age

Article Title: Revealing system-level correlations between aging and calorie restriction using a mouse transcriptome

doi: 10.1007/s11357-009-9106-3

Figure Lengend Snippet: Over-represented GO terminologies in the aging and CR transcriptomes ( BP biological process, CC cellular component)

Article Snippet: The major compartments are displayed on the inner bar graph table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Process GO context GO description Up Down Fractions (%) Fold change c P value d Aging/CR a Whole b Aging BP Muscle development 9 9 5.76% 0.88% 6.52 6.38 × 10 -23 Muscle contraction 4 7 3.75% 0.51% 7.37 7.62 × 10 -18 Cell adhesion 21 8 8.36% 3.88% 2.15 1.21 × 10 -5 Nervous system development 9 4 3.75% 1.28% 2.92 3.55 × 10 -5 Cellular protein metabolism 45 30 21.61% 14.30% 1.51 7.76 × 10 -5 Lipid metabolism 10 3.46% 1.25% 2.77 0.0001 Immune response 16 4 5.76% 3.13% 1.84 0.0041 Amino acid metabolism 3 5 2.31% 1.06% 2.18 0.0212 Cell growth 3 1.44% 0.57% 2.54 0.0277 Cell-cell signaling 6 4 3.17% 1.70% 1.86 0.0316 Cytoskeleton organization 7 6 4.32% 2.58% 1.67 0.0383 CC Contractile fiber 5 4 2.93% 0.49% 5.99 5.66 × 10 -11 Mitochondrion 24 22 13.49% 5.80% 2.32 7.52 × 10 -10 Extracellular space 54 37 26.69% 15.98% 1.67 4.53 × 10 -5 Lysosome 8 3 3.23% 0.88% 3.65 2.75 × 10 -6 Peroxisome 0 6 1.76% 0.52% 3.37 0.001296 CR BP Immune response 6 29 7.81% 3.13% 2.50 6.26 × 10 -9 Lipid metabolism 11 6 3.79% 1.25% 3.03 7.84 × 10 -7 Response to stimulus 11 29 9.15% 4.71% 1.94 6.09 × 10 -6 Cell proliferation 8 16 5.36% 2.34% 2.29 1.71 × 10 -5 Glucose catabolism 4 3 1.56% 0.39% 3.99 5.28 × 10 -5 Cholesterol metabolism 5 1.56% 0.40% 3.90 7.13 × 10 -5 Angiogenesis 3 6 2.01% 0.63% 3.21 0.0001 Cell adhesion 7 24 6.92% 3.88% 1.78 0.0007 Cell cycle 10 23 7.37% 4.38% 1.68 0.0016 Electron transport 13 10 5.13% 2.76% 1.86 0.0017 Muscle development 7 2.01% 0.88% 2.27 0.0094 Cytoskeleton organization 3 17 4.46% 2.58% 1.73 0.0105 Chemotaxis 0 7 1.56% 0.68% 2.31 0.0194 Amino acid metabolism 5 4 2.01% 1.06% 1.90 0.0451 CC Extracellular space 38 84 27.29% 15.98% 1.71 3.02 × 10 -11 Lysosome 3 11 3.13% 0.88% 3.55 2.27 × 10 -7 Mitochondrion 31 19 11.41% 5.80% 1.97 2.44 × 10 -7 Endoplasmic reticulum 13 22 8.05% 3.78% 2.13 1.41 × 10 -6 Open in a separate window a The ratio for the DEGs involved in the terminology among the aging/CR transcriptomes b The ratio for the DEGs involved in the terminology among the overall mouse transcriptomes c The ratio between the aging/CR fraction and the whole fraction d P values < 0.05 are considered significant Over-represented GO terminologies in the aging and CR transcriptomes ( BP biological process, CC cellular component) Interestingly, the genes related to cell adhesion and immune responses were found mostly up-regulated with aging.

Techniques: Chemotaxis Assay

Journal: Age

Article Title: Revealing system-level correlations between aging and calorie restriction using a mouse transcriptome

doi: 10.1007/s11357-009-9106-3

Figure Lengend Snippet: Transcription factors showing the consistent expression patterns with aging- or CR-related genes

Article Snippet: The major compartments are displayed on the inner bar graph table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Process GO context GO description Up Down Fractions (%) Fold change c P value d Aging/CR a Whole b Aging BP Muscle development 9 9 5.76% 0.88% 6.52 6.38 × 10 -23 Muscle contraction 4 7 3.75% 0.51% 7.37 7.62 × 10 -18 Cell adhesion 21 8 8.36% 3.88% 2.15 1.21 × 10 -5 Nervous system development 9 4 3.75% 1.28% 2.92 3.55 × 10 -5 Cellular protein metabolism 45 30 21.61% 14.30% 1.51 7.76 × 10 -5 Lipid metabolism 10 3.46% 1.25% 2.77 0.0001 Immune response 16 4 5.76% 3.13% 1.84 0.0041 Amino acid metabolism 3 5 2.31% 1.06% 2.18 0.0212 Cell growth 3 1.44% 0.57% 2.54 0.0277 Cell-cell signaling 6 4 3.17% 1.70% 1.86 0.0316 Cytoskeleton organization 7 6 4.32% 2.58% 1.67 0.0383 CC Contractile fiber 5 4 2.93% 0.49% 5.99 5.66 × 10 -11 Mitochondrion 24 22 13.49% 5.80% 2.32 7.52 × 10 -10 Extracellular space 54 37 26.69% 15.98% 1.67 4.53 × 10 -5 Lysosome 8 3 3.23% 0.88% 3.65 2.75 × 10 -6 Peroxisome 0 6 1.76% 0.52% 3.37 0.001296 CR BP Immune response 6 29 7.81% 3.13% 2.50 6.26 × 10 -9 Lipid metabolism 11 6 3.79% 1.25% 3.03 7.84 × 10 -7 Response to stimulus 11 29 9.15% 4.71% 1.94 6.09 × 10 -6 Cell proliferation 8 16 5.36% 2.34% 2.29 1.71 × 10 -5 Glucose catabolism 4 3 1.56% 0.39% 3.99 5.28 × 10 -5 Cholesterol metabolism 5 1.56% 0.40% 3.90 7.13 × 10 -5 Angiogenesis 3 6 2.01% 0.63% 3.21 0.0001 Cell adhesion 7 24 6.92% 3.88% 1.78 0.0007 Cell cycle 10 23 7.37% 4.38% 1.68 0.0016 Electron transport 13 10 5.13% 2.76% 1.86 0.0017 Muscle development 7 2.01% 0.88% 2.27 0.0094 Cytoskeleton organization 3 17 4.46% 2.58% 1.73 0.0105 Chemotaxis 0 7 1.56% 0.68% 2.31 0.0194 Amino acid metabolism 5 4 2.01% 1.06% 1.90 0.0451 CC Extracellular space 38 84 27.29% 15.98% 1.71 3.02 × 10 -11 Lysosome 3 11 3.13% 0.88% 3.55 2.27 × 10 -7 Mitochondrion 31 19 11.41% 5.80% 1.97 2.44 × 10 -7 Endoplasmic reticulum 13 22 8.05% 3.78% 2.13 1.41 × 10 -6 Open in a separate window a The ratio for the DEGs involved in the terminology among the aging/CR transcriptomes b The ratio for the DEGs involved in the terminology among the overall mouse transcriptomes c The ratio between the aging/CR fraction and the whole fraction d P values < 0.05 are considered significant Over-represented GO terminologies in the aging and CR transcriptomes ( BP biological process, CC cellular component) Interestingly, the genes related to cell adhesion and immune responses were found mostly up-regulated with aging.

Techniques: Expressing, Transduction, Ubiquitin Proteomics

Journal: Age

Article Title: Revealing system-level correlations between aging and calorie restriction using a mouse transcriptome

doi: 10.1007/s11357-009-9106-3

Figure Lengend Snippet: Functional associations at system levels. Systems in the aging and CR transcriptomes are closely correlated in a positive (red line) or negative (blue line) manner. Lipid metabolism, especially, showed an inverse correlation with many other systems, including immune response, cell cycle, cell proliferation, and muscle development. The systems showing the strongest inverse correlation were lipid metabolism and immune response (P = 6.73 × 10-9). The relationship between lipid metabolism and cytoskeleton organization is somewhat ambiguous (green line), showing positive and negative patterns together

Article Snippet: The major compartments are displayed on the inner bar graph table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Process GO context GO description Up Down Fractions (%) Fold change c P value d Aging/CR a Whole b Aging BP Muscle development 9 9 5.76% 0.88% 6.52 6.38 × 10 -23 Muscle contraction 4 7 3.75% 0.51% 7.37 7.62 × 10 -18 Cell adhesion 21 8 8.36% 3.88% 2.15 1.21 × 10 -5 Nervous system development 9 4 3.75% 1.28% 2.92 3.55 × 10 -5 Cellular protein metabolism 45 30 21.61% 14.30% 1.51 7.76 × 10 -5 Lipid metabolism 10 3.46% 1.25% 2.77 0.0001 Immune response 16 4 5.76% 3.13% 1.84 0.0041 Amino acid metabolism 3 5 2.31% 1.06% 2.18 0.0212 Cell growth 3 1.44% 0.57% 2.54 0.0277 Cell-cell signaling 6 4 3.17% 1.70% 1.86 0.0316 Cytoskeleton organization 7 6 4.32% 2.58% 1.67 0.0383 CC Contractile fiber 5 4 2.93% 0.49% 5.99 5.66 × 10 -11 Mitochondrion 24 22 13.49% 5.80% 2.32 7.52 × 10 -10 Extracellular space 54 37 26.69% 15.98% 1.67 4.53 × 10 -5 Lysosome 8 3 3.23% 0.88% 3.65 2.75 × 10 -6 Peroxisome 0 6 1.76% 0.52% 3.37 0.001296 CR BP Immune response 6 29 7.81% 3.13% 2.50 6.26 × 10 -9 Lipid metabolism 11 6 3.79% 1.25% 3.03 7.84 × 10 -7 Response to stimulus 11 29 9.15% 4.71% 1.94 6.09 × 10 -6 Cell proliferation 8 16 5.36% 2.34% 2.29 1.71 × 10 -5 Glucose catabolism 4 3 1.56% 0.39% 3.99 5.28 × 10 -5 Cholesterol metabolism 5 1.56% 0.40% 3.90 7.13 × 10 -5 Angiogenesis 3 6 2.01% 0.63% 3.21 0.0001 Cell adhesion 7 24 6.92% 3.88% 1.78 0.0007 Cell cycle 10 23 7.37% 4.38% 1.68 0.0016 Electron transport 13 10 5.13% 2.76% 1.86 0.0017 Muscle development 7 2.01% 0.88% 2.27 0.0094 Cytoskeleton organization 3 17 4.46% 2.58% 1.73 0.0105 Chemotaxis 0 7 1.56% 0.68% 2.31 0.0194 Amino acid metabolism 5 4 2.01% 1.06% 1.90 0.0451 CC Extracellular space 38 84 27.29% 15.98% 1.71 3.02 × 10 -11 Lysosome 3 11 3.13% 0.88% 3.55 2.27 × 10 -7 Mitochondrion 31 19 11.41% 5.80% 1.97 2.44 × 10 -7 Endoplasmic reticulum 13 22 8.05% 3.78% 2.13 1.41 × 10 -6 Open in a separate window a The ratio for the DEGs involved in the terminology among the aging/CR transcriptomes b The ratio for the DEGs involved in the terminology among the overall mouse transcriptomes c The ratio between the aging/CR fraction and the whole fraction d P values < 0.05 are considered significant Over-represented GO terminologies in the aging and CR transcriptomes ( BP biological process, CC cellular component) Interestingly, the genes related to cell adhesion and immune responses were found mostly up-regulated with aging.

Techniques: Functional Assay

Journal: EBioMedicine

Article Title: CD24-targeted intraoperative fluorescence image-guided surgery leads to improved cytoreduction of ovarian cancer in a preclinical orthotopic surgical model.

doi: 10.1016/j.ebiom.2020.102783

Figure Lengend Snippet: Fig. 1. CD24 is highly expressed in ovarian cancer cell lines and primary ovarian cancer cells, and can be detected by CD24-AF750 NIR staining. (a) Flow cytometry analysis of EOC cell lines OV-90, Caov-3, COV318 and Skov-3 stained with the monoclonal antibody CD24 conjugated to AF750 (n = 3). Each column represents the mean with SD. (b) CD24 expres- sion was evaluated by flow cytometry, and antigen-binding capacity was quantified for OV-90 compared to monoclonal IgG1 antibody (mAb) control. (c) Validation of target-specific binding. White light (colour) image, near infrared (NIR 800) fluorescent image and pseudo-coloured fluorescence intensity merge image of CD24-AF750 stained EOC cell pellets (25 £ 107 tumour cells, scale bar = 1 cm). (d) Immunohistochemistry of four different patient-derived xenograft (PDX) ovarian tumours, stained with anti-CD24. Immunoreactivity score (IRS) 425 § 096 (scale: 1ow = 1 7 = high). Images were captured at 100£ and 200£ magnification (scale bar = 25 mm).

Article Snippet: An unconjugated monoclonal CD33 IgG1 antibody (Clone WM53, Cat# MCA1271T, RRID: AB_2074234, Bio-Rad) was used as a negative control.

Techniques: Staining, Flow Cytometry, Cytometry, Binding Assay, Control, Biomarker Discovery, Immunohistochemistry, Derivative Assay