Journal: bioRxiv

Article Title: A bivalent lysine-acetylated small-molecule binding site in MYC

doi: 10.64898/2026.03.02.707789

Figure Lengend Snippet: (A) Representative FISH image of PC3M cells. The MYC dual color breakapart probe (Abbott Park, Illinois) labeled the 5’MYC as red and the 3’MYC as green. The normal signal pattern shows two fusions (overlap with the green and red signals). Single green and/or red signals indicate MYC rearrangements. Nuclei are counter-stained with DAPI (blue). (B) Colony-formation assays were performed comparing PC3-MCas9 cells to PC3-MCas9 lines co trans-fected with pairs of enriched sgRNA guides: (A169/A361) and (A169/ A423) following 14 days of treatment with 6μM MYCi975. Data was normalized to DMSO control and represent the mean ± S.D. from biological replicates. (*p<0.05; **p<0.01; ***p<0.001). (C) Relative cell viability of PC3-MCas9 cells co-transfected with sgRNA pairs (A11/A362, A169/A423, and A11/A423) was measured following continuous exposure to MYCi975 at 6µM for 20 days, followed by 10µM until day 35. (D, E) Multiple-sequence alignment of the MYC emBII and bHLH-LZ regions across vertebrate species. Blue dots denote DNA-binding residues in BHLH; green dots mark leucine-zipper dimerization residues. The MYC mutations identified are indicated in red font. (F) Expression and BLI K d of recombinant MYC mutants WAL-QRA, WAL (emBII mutant), and QRA (bHLH mutant) with MYCi975. (G) Colony formation assay of HO15.19 fibroblast, 22Rv1 and PC3 cells stably expressing Flag-tagged WT MYC and WAL-QRA MYC after treatment with DMSO and varying concentrations of MYCi975 for 14 days. Representative wells of colony formation assay are shown. (H) Cell viability of HO15.19 MYC and WAL-QRA MYC mutant was measured by IncuCyte live-cell imag-ing for 4 days with varying concentrations of MYCi975.

Article Snippet: 22Rv1, PC3, MyC-CaP, HEK293, MV-4-11, PC12, A549, LLC1 and HL-60 cell lines were purchased from the American Type Culture Collection (ATCC).

Techniques: Labeling, Staining, Control, Transfection, Sequencing, Binding Assay, Expressing, Recombinant, Mutagenesis, Colony Assay, Stable Transfection

Journal: bioRxiv

Article Title: A bivalent lysine-acetylated small-molecule binding site in MYC

doi: 10.64898/2026.03.02.707789

Figure Lengend Snippet: (A) Schematic of the CRISPR Cas9 suppressor screening strategy. A Cas9-expressing PC-3M cell pool was transduced with a pooled sgRNA library targeting the MYC locus. Following enrichment in 6 µM MYCi975, resistant clones were isolated and sequenced to identify mutations conferring MYCi975 resistance. (B) Volcano plot summarizing the enrichment of sgRNAs guides across two independent screens. sgRNA guides targeting residues A169, A361, and A423 were significantly enriched. Deep sequencing of resistant clones identified mutation hotspots clustering close to the eMBII region (amino acid residues 196–200) and the bHLH (amino acid residues 370–380) domains of MYC. Representative sequences are shown. (C) BLI K d of MYCi975 for recombinant WT MYC, WAL-QRA MYC mutants, and individual domain MYC mutants (WAL and QRA). (D) Quantification of colony formation assays in HO15.19 fibroblast, 22Rv1 and PC3 cells expressing Flag-tagged WT MYC and WAL-QRA MYC mutant after MYCi975 treatment for 14 days. Data was normalized to DMSO control and represent the mean ± S.D. from biological replicates. *Denote statistical significance (*p<0.05; **p<0.01; ***p<0.001). (E) Immunoblot analyses of FLAG-tagged MYC protein stability in HO15.19, 22Rv1 and PC3 cells express-ing Flag-MYC WT or Flag-WAL-QRA-MYC mutant, treated with 6µM MYCi975 for 24 hours. GAPDH serves as a loading control.

Article Snippet: 22Rv1, PC3, MyC-CaP, HEK293, MV-4-11, PC12, A549, LLC1 and HL-60 cell lines were purchased from the American Type Culture Collection (ATCC).

Techniques: CRISPR, Expressing, Transduction, Clone Assay, Isolation, Sequencing, Mutagenesis, Recombinant, Control, Western Blot

Journal: bioRxiv

Article Title: A bivalent lysine-acetylated small-molecule binding site in MYC

doi: 10.64898/2026.03.02.707789

Figure Lengend Snippet: (A) We analyzed high-resolution acetylome data from the Clinical Proteomic Tumor Analysis Consortium (CPTAC), including 8 cohorts totaling 928 tumors and 389 normal tissues. K148 MYC modification was significantly elevated in tumors compared to matched solid normal tissues in 5 of 8 cohorts examined: breast cancer (PDC000239; p<0.0001), lung squamous cell carcinoma (PDC000233; p<0.0001), lung adenocarci-noma (PDC000491; p=0.0184), glioblastoma (PDC000245; p=0.028), and a multi-cancer cohort (PDC000450; p=0.009) comprising breast cancer, epithelial and complex epithelial neoplasms, glioblastoma, lung cancer, meningiomas, skin cancer and uterine adenocarcinoma. One lung cancer cohort (PDC000224) and two uterine corpus endometrial carcinoma cohorts (PDC000226, PDC000443) did not follow this pattern. Each data point represents an individual patient sample. Statistical analysis was performed using a two-tailed Mann-Whitney U test. p-values: ****p<0.0001, *p<0.05. (Abbreviations: AM, Additional Metastatic; M, Metastatic; PT, Primary Tumor; RT, Recurrent Tumor; N, Normal). (B) Unsupervised clustering of MYC target genes based on responses to wild-type MYC versus acetylation-deficient MYC (K148R) identified four clusters comprising MYC-repressed (clusters 1, 4) and MYC-induced (clusters 2, 3) programs. MYCi975 selectively induced acetylation-dependent stress-response genes (cluster 1) and repressed acetylation-dependent canonical MYC targets (cluster 2), while minimally affecting acety-lation-independent programs (clusters 3, 4), indicating preferential disruption of acetylated MYC dependent transcription. (C) Functional enrichment analysis of each gene cluster highlighting the top 10 significantly enriched path-ways. (D) Inverse correlation between AcK148-MYC abundance and sensitivity to MYCi975. Scatter plot showing the inverse relationship between the relative abundance of the AcK148-MYC modification (AcK148-MYC:MYC ratio) and cellular sensitivity to the MYC inhibitor MYCi975 (IC50) across a panel of 12 cancer cell lines including leukemia (Kasumi-1, HL-60, THP-1, MV-4:11), Prostate (PC3 and 22Rv1), Liver (PLC/PRF/5, HepG2), lung (A549), breast (MDA-MB-231), ovarian (A2780) and colorectal (HT-29) cell lines. A higher ratio of AcK148-MYC is associated with a lower IC50, indicating increased sensitivity to MYCi975.

Article Snippet: 22Rv1, PC3, MyC-CaP, HEK293, MV-4-11, PC12, A549, LLC1 and HL-60 cell lines were purchased from the American Type Culture Collection (ATCC).

Techniques: Modification, Two Tailed Test, MANN-WHITNEY, Disruption, Functional Assay

Journal: bioRxiv

Article Title: A bivalent lysine-acetylated small-molecule binding site in MYC

doi: 10.64898/2026.03.02.707789

Figure Lengend Snippet: (A) Chemical structure of new MYCI analog MYCi648. (B) Binding affinities K d of MYC inhibitors to WT MYC and acetylated MYC (AcK148/157-MYC), along with cellular activity measured by E-box luciferase inhibition, CMV luciferase control assays, and viability assays in PC3 and control PC12 cells. (C) Tumor growth inhibition (TGI) in the MycCaP/FVB/N prostate cancer model treated with vehicle (n=8) or MYCi648 (30 mg/kg/d; n=5) i.p. (***p<0.001). (D) Tumor growth inhibition in the LLC1 syngeneic model (C57BL/6 mice) treated with vehicle (n=6) and MYCi648 (50 mg/kg/d; n=6) i.p. (***p<0.001). (E) BLI K d of MYCi648 to WT MYC and the WAL-QRA MYC mutant, demonstrating reduced binding to the WAL-QRA MYC mutant protein. (F) Quantification of colony formation assays in 22Rv1 and PC3 cells expressing WT MYC and WAL-QRA MYC mutant after MYCi648 treatment for 14 days. Data was normalized to DMSO control and represent the mean ± S.D. from biological replicates. *Denote statistical significance (*p<0.05; **p<0.01; ***p<0.001).

Article Snippet: 22Rv1, PC3, MyC-CaP, HEK293, MV-4-11, PC12, A549, LLC1 and HL-60 cell lines were purchased from the American Type Culture Collection (ATCC).

Techniques: Binding Assay, Activity Assay, Luciferase, Inhibition, Control, Mutagenesis, Expressing

Journal: EMBO reports

Article Title: eIF5A is required for autophagy by mediating ATG3 translation.

doi: 10.15252/embr.201846072

Figure Lengend Snippet: Figure 4. eIF5A regulates translation of ATG3.

Article Snippet: Primary antibodies used are as follows: LC3B (nanotools, 1:200) (LC3B CST, 1:1,000), GABARAP (Abgent 1:1,000), GATE-16 (MBL, 1:1,000), ATG3 (Sigma, 1:500), Vinculin (Sigma, 1:100,000), GAPDH (Santacruz, 1:20,000), eIF5A (Santacruz, 1:500), Lamin A1 (Santacruz, 1:1,000), histone H3 (Abcam 1:10,000), p62 (MBL, 1:5,000), RFP/ Cherry (Rockland, 1:6,000), p-mTOR (Ser 2448) (CST, 1:1,000), mTOR (Cell signaling, 1:1,000), RPL23A (Abcam, 1:50,000), RPS6 (CST, 1:1,000), RPL10A (Santacruz, 1:2,000), Hypusine (Merck Millipore 1:5,000).

Techniques:

Journal: EMBO reports

Article Title: eIF5A is required for autophagy by mediating ATG3 translation.

doi: 10.15252/embr.201846072

Figure Lengend Snippet: Figure 5. Regulation of autophagy by eIF5A is mediated via ATG3 translation.

Article Snippet: Primary antibodies used are as follows: LC3B (nanotools, 1:200) (LC3B CST, 1:1,000), GABARAP (Abgent 1:1,000), GATE-16 (MBL, 1:1,000), ATG3 (Sigma, 1:500), Vinculin (Sigma, 1:100,000), GAPDH (Santacruz, 1:20,000), eIF5A (Santacruz, 1:500), Lamin A1 (Santacruz, 1:1,000), histone H3 (Abcam 1:10,000), p62 (MBL, 1:5,000), RFP/ Cherry (Rockland, 1:6,000), p-mTOR (Ser 2448) (CST, 1:1,000), mTOR (Cell signaling, 1:1,000), RPL23A (Abcam, 1:50,000), RPS6 (CST, 1:1,000), RPL10A (Santacruz, 1:2,000), Hypusine (Merck Millipore 1:5,000).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Phosphorylation of the oncogenic transcription factor ERG in prostate cells dissociates polycomb repressive complex 2, allowing target gene activation

doi: 10.1074/jbc.M117.796458

Figure Lengend Snippet: Phosphorylation of ERG at Ser-96 decreases its affinity for the PRC2 complex. A, pulldown of EZH2/CBP/EWS from PC3 nuclear extracts. The indicated N-terminal His-tagged ERG constructs were immobilized to beads, incubated with PC3 nuclear extract, and washed. EZH2/CBP/EWS binding was visualized by immunoblot. Ponceau stain of the same blots show input amount of His-ERG. B, pulldown of EZH2 as in A but with full-length and indicated deletion mutants of ERG. EZH2/CBP/EWS binding was visualized by immunoblot (top panels, IP). Ponceau stain shows amount of His-ERG present (bottom panels, Input). C, co-immunoprecipitation using ERG antibody from nuclear extracts prepared from RWPE1 cells expressing vector or indicated FLAG-tagged WT and mutants of ERG, followed by immunoblotting for EZH2, SUZ12, and FLAG. D, co-immunoprecipitation of ERG as in C but with cells treated either with 20 μm U0126 or mock treatment for 6 h. E, VCaP cells with or without PMA treatment (1 h 200 nm) were immunoprecipitated with ERG antibody or IgG control and immunoblotted for indicated proteins.

Article Snippet: To the bead-bound ETS proteins, ∼14 μg of PC3 nuclear extracts (catalog no. sc-2152; Santa Cruz Biotechnology) were added and rotated for 1–2 h. The beads were washed four times with Nonidet P-40 lysis buffer, and bead-bound proteins were eluted using SDS protein loading dye.

Techniques: Phospho-proteomics, Construct, Incubation, Binding Assay, Western Blot, Staining, Immunoprecipitation, Expressing, Plasmid Preparation, Control

Journal: PLoS ONE

Article Title: Cysteine (C)-X-C Receptor 4 Undergoes Transportin 1-Dependent Nuclear Localization and Remains Functional at the Nucleus of Metastatic Prostate Cancer Cells

doi: 10.1371/journal.pone.0057194

Figure Lengend Snippet: A , A human prostate tissue array, ranging from normal to high-grade prostate cancer, was evaluated by IHC for CXCR4 expression using standard methods. Samples were evaluated at magnification 40X, using a Q-Imaging camera of Olympus BX51 Microscope with Bioquant® Image Analysis Software (RtmBometrics). Normal prostate tissues demonstrated slightly weak or undetectable brown staining for CXCR4 (positive cells<5%), and no CXCR4 expression in the nucleus. Representative low grade prostate tissue (grade 2, stage II, T 2 N 0 M 0 , adenocarcinoma) demonstrated random/focal positive staining for CXCR4 in the nucleus (positive cells >11%, but less than 50%), indicating low expression of CXCR4. Representative high grade metastatic prostate tissue (grade 4, stage IV, T 4 N 1 M 1 , adenocarcinoma) demonstrated diffuse/intense staining (positive cells >50%), indicating high expression for CXCR4 in the nucleus. Scale bar represents 50 µm. B , CXCR4 IgG2B mouse monoclonal antibody was evaluated for specificity to CXCR4 protein by western blot analysis in PC3 (CXCR4 positive) or 293T (CXCR4 null) cell lines. C , CXCR4 antibody was evaluated for specificity to CXCR4 protein by immunoprecipitation for CXCR4 and western blot analysis for CXCR4. D , CXCR4 IgG2B antibody was evaluated for specificity to CXCR4 protein by immunoprecipitation with Fibronectin IgG2B mouse monoclonal antibody (unrelated isotype control) and western blot analysis for CXCR4; expression of Fibronectin protein was confirmed by western blot analysis. Beta-actin was used as a loading control.

Article Snippet: One milligram of PC3 whole cell lysates were immunoprecipitated for CXCR4 (Santa Cruz; 1 μg per 250 μg of protein) overnight at 4°C, followed by incubation with Protein A/G Plus-Agarose beads (Santa Cruz) for 2 hrs at 4°C.

Techniques: Expressing, Imaging, Microscopy, Software, Staining, Western Blot, Immunoprecipitation, Control

Journal: PLoS ONE

Article Title: Cysteine (C)-X-C Receptor 4 Undergoes Transportin 1-Dependent Nuclear Localization and Remains Functional at the Nucleus of Metastatic Prostate Cancer Cells

doi: 10.1371/journal.pone.0057194

Figure Lengend Snippet: A , Normal prostate epithelial (RWPE1) and PCa (PC3, DU145, 22RV1) cells were stimulated with SDF1α (100 ng/ µl) prior to subcellular fractionation into non-nuclear and nuclear fractions. Immunoblots were probed with anti-CXCR4. Anti-CD44 (non-nuclear) and anti-Topoisomerase1 (Topo 1, nuclear) were used as markers for fractionation purity and as loading controls. The bar graphs are quantitative results of the band density representing expression of CXCR4 in each fraction. Data were mean + S.E. from three independent experiments. *, P<0.05. B , Immunocytochemistry of PCa cell lines for CXCR4. PCa cells were stimulated with SDF1α (100 ng/ µl), fixed with methanol, blocked then incubated with an antibody mixture containing a mouse anti-CXCR4 monoclonal antibody and a rabbit polyclonal anti-Lamin A/C antibody, followed by secondary mixture containing a Cy3-conjugated anti-mouse antibody and FITC-conjugated anti-rabbit antibody. Imaging was with a Zeiss LSM-510 UV Confocal Microscope using the 63× Plan-Apochromat 63x/1.40 Oil DIC objective at excitation 488 nm for FITC and 543 nm for Cy3. Confocal images demonstrating the plasma membrane and cytosolic localization of CXCR4 (red), intact nuclear membrane (green), and nuclear-associated localization of CXCR4 (yellow/orange) are shown. Small arrows indicate co-localization of CXCR4 with the nucleus (yellow/orange). Scale bars represent 50 µm.

Article Snippet: One milligram of PC3 whole cell lysates were immunoprecipitated for CXCR4 (Santa Cruz; 1 μg per 250 μg of protein) overnight at 4°C, followed by incubation with Protein A/G Plus-Agarose beads (Santa Cruz) for 2 hrs at 4°C.

Techniques: Fractionation, Western Blot, Expressing, Immunocytochemistry, Incubation, Imaging, Microscopy, Clinical Proteomics, Membrane

Journal: PLoS ONE

Article Title: Cysteine (C)-X-C Receptor 4 Undergoes Transportin 1-Dependent Nuclear Localization and Remains Functional at the Nucleus of Metastatic Prostate Cancer Cells

doi: 10.1371/journal.pone.0057194

Figure Lengend Snippet: A , GFP-CXCR4 fusion protein localized similar to endogenous CXCR4. CXCR4-pEGFPN1 transfected PC3 cells were stimulated with SDF1α, fixed with methanol, blocked then incubated with a mouse anti-CXCR4 monoclonal antibody, followed by a Cy3-conjugated anti-mouse secondary antibody. Nuclei were stained with DAPI (blue). Images were taken at 40× maginification using Axiovision software 4.8.2 with a Zeiss Axio Imager.z1 fluorescence microscope at ex = 470 nm for FITC, ex = 358 nm for DAPI and ex = 551 nm for Cy3. Images demonstrate the co-localization (yellow) of endogenous CXCR4 (red) with GFP-tagged CXCR4 (green). B , Localization analysis of wild type CXCR4 (CXCR4-pEGFPN1), NLS-mutant of CXCR4 (pEGFPN1-CXCR4 R146A ,) and deleted NLS of CXCR4 (CXCR4 ΔNLS ) by immunocytochemistry in PC3 cells. Nuclei were stained with propidium iodide (red) and CXCR4 was detected as the fusion protein GFP-CXCR4 (green). Imaging was with a Zeiss LSM-510 UV Confocal Microscope using the 63× Plan-Apochromat 63x/1.40 Oil DIC objective at ex = 488 nm for FITC and ex = 543 nm for Cy3. Scale bars represent 50 µm. C , Transfected cells were stimulated with SDF1α prior to subcellular fractionation into non-nuclear and nuclear fractions. Immunoblots were probed with anti-GFP to detect the fusion protein GFP-CXCR4. Anti-CD44 (non-nuclear) and anti-Topoisomerase1 (Topo 1, nuclear) were used as markers for fractionation purity and as loading controls.

Article Snippet: One milligram of PC3 whole cell lysates were immunoprecipitated for CXCR4 (Santa Cruz; 1 μg per 250 μg of protein) overnight at 4°C, followed by incubation with Protein A/G Plus-Agarose beads (Santa Cruz) for 2 hrs at 4°C.

Techniques: Transfection, Incubation, Staining, Software, Fluorescence, Microscopy, Mutagenesis, Immunocytochemistry, Imaging, Fractionation, Western Blot

Journal: PLoS ONE

Article Title: Cysteine (C)-X-C Receptor 4 Undergoes Transportin 1-Dependent Nuclear Localization and Remains Functional at the Nucleus of Metastatic Prostate Cancer Cells

doi: 10.1371/journal.pone.0057194

Figure Lengend Snippet: A , Sixty micrograms of total protein were analyzed for TRN1 expression by western blot analysis using a TRN1 specific antibody. Alpha-tubulin served as a loading control. B , One milligram of PC3 whole cell lysate was immunoprecipitated with anti-CXCR4 and separated by SDS-PAGE. Immunocomplexes were probed with anti-TRN1 or anti-CXCR4 to ensure that CXCR4 interacted with TRN1 and was immunoprecipitated, respectively. Thirty micrograms of whole cell PC3 supernatant, post-immunoprecipitation, were separated by SDS-PAGE and harvested for western blot analysis to assess the efficiency of CXCR4 immunoprecipitation. C and D , Cells were transiently transfected with TRN1-specific siRNA to determine an effective concentration ( C ) , prior to harvesting for immunohistochemistry with anti-Lamin A/C and anti-CXCR4 ( D ) . Images were taken using Zeiss Axio Imager.z1 fluorescence microscope at 40× magnification at excitation 470 nm for FITC and 551 nm for Cy3. Small arrows indicate co-localization of CXCR4 with the nucleus (yellow/orange). Scale bar represents 50 µm.

Article Snippet: One milligram of PC3 whole cell lysates were immunoprecipitated for CXCR4 (Santa Cruz; 1 μg per 250 μg of protein) overnight at 4°C, followed by incubation with Protein A/G Plus-Agarose beads (Santa Cruz) for 2 hrs at 4°C.

Techniques: Expressing, Western Blot, Control, Immunoprecipitation, SDS Page, Transfection, Concentration Assay, Immunohistochemistry, Fluorescence, Microscopy

Journal: PLoS ONE

Article Title: Cysteine (C)-X-C Receptor 4 Undergoes Transportin 1-Dependent Nuclear Localization and Remains Functional at the Nucleus of Metastatic Prostate Cancer Cells

doi: 10.1371/journal.pone.0057194

Figure Lengend Snippet: A , Representative light images of whole cells and isolated nuclei confirmed the integrity of nuclear isolation at 20× magnification. B , Whole cells were treated with SDF1α prior to isolating and lysing intact nuclei. Nuclei lysates (1 mg) were immunoprecipitated with anti-CXCR4 and separated by SDS-PAGE. Immunocomplexes were probed for G αi (first row) or CXCR4 antibody (second row), respectively. Anti-CD44 (non-nuclear) and anti-Topoisomerase1 (Topo1, nuclear) were used as markers for fractionation purity and as loading controls. C , PC3 nuclei were isolated, incubated with FluoForte dye Ca 2+ probe, followed by incubation with AMD3100 or pertussis toxin (PTX) for 1 hr, then stimulated with SDF1α for 30 min. An increase in fluorescent-bound Ca 2+ was measured on a microplate reader at ex = 490 nm/em = 525 nm.

Article Snippet: One milligram of PC3 whole cell lysates were immunoprecipitated for CXCR4 (Santa Cruz; 1 μg per 250 μg of protein) overnight at 4°C, followed by incubation with Protein A/G Plus-Agarose beads (Santa Cruz) for 2 hrs at 4°C.

Techniques: Isolation, Immunoprecipitation, SDS Page, Fractionation, Incubation

Journal: Bioactive Carbohydrates and Dietary Fibre

Article Title: Resistant starch from a tuberous root from the Andes cordillera improves metabolic and immune parameters in broilers.

doi: 10.1016/j.bcdf.2024.100420

Figure Lengend Snippet: Fig. 3. CD3+, CD4+ and CD8+ intestinal T lymphocytes subpopulations in broilers on day 35 determined by flow cytometry (upper left image). The images A, B, C correspond to dot plot made in the FlowJo XV software based on readings made in the Facs Canto II flow cytometer (Becton Dickinson). The images show: A. Distribution by size (FSC-A) and complexity (SSC-A) of the events analyzed in broilers, B. Percentage of CD4+ T cells in broiler chickens fed with a control diet C. Percentage of CD4+ T cells in broiler chickens fed with a diet supplemented with RS2.

Article Snippet: The labeling mix 2 was prepared with 2 μL of each of the following anti-chicken monoclonal antibodies: anti-CD3 epsilon (PC3/188A), antiCD8 alpha (EP72), the anti-human antibody anti-MHC1 (F21-2) coupled to FITC (Novus biologicals, Colorado, USA), which was used as a positive control, and 94 μL of cytometer buffer.

Techniques: Flow Cytometry, Software, Control

Journal: Frontiers in immunology

Article Title: Investigating mammary glands of lactating goats for the presence of tertiary lymphoid organs.

doi: 10.3389/fimmu.2022.941333

Figure Lengend Snippet: FIGURE 1 Representative images of hematoxylin and eosin staining (HE) and immunohistochemistry against CD20 (B cells), CD3 (T cells), MECA79 (high endothelial venules; HEVs), CD40 (follicular dendritic cells), and BCL6 (germinal center) in a small-sized aggregation of lymphocytes in goat mammary gland tissues. Scale bar, 100 mm or 25 mm (clippings).

Article Snippet: The sections were then incubated overnight at 4°C with either rabbit polyclonal antibodies against goat-IgA (#A50-106A, Bethyl Laboratories), claudin-3 (#34-1700, Thermo Fisher Scientific), claudin-4 (#PA5-32354, Thermo Fisher Scientific), or mouse monoclonal antibodies against BCL6 (#sc7388, Santa Cruz Biotechnology), CD3 (#NBP2-53386, Novus Biologicals), CD20 (#NBP2-45454, Novus Biologicals), CD40 (#sc-13128, Santa Cruz Biotechnology), occludin (#sc-133256, Santa Cruz Biotechnology), or a rat monoclonal antibody against MECA79 (#sc-19602, Santa Cruz Biotechnology) diluted in PBS-T containing 2.5% bovine serum albumin.

Techniques: Staining, Immunohistochemistry

Journal: Frontiers in immunology

Article Title: Investigating mammary glands of lactating goats for the presence of tertiary lymphoid organs.

doi: 10.3389/fimmu.2022.941333

Figure Lengend Snippet: FIGURE 2 Representative images of hematoxylin and eosin staining (HE) staining and immunohistochemistry against CD20 (B cells), CD3 (T cells), MECA79 (high endothelial venules; HEVs), CD40 (follicular dendritic cells), and BCL6 (germinal center) in a large-sized aggregation of lymphocytes in goat mammary glands. Images of the same position are shown. Scale bar, 100 mm.

Article Snippet: The sections were then incubated overnight at 4°C with either rabbit polyclonal antibodies against goat-IgA (#A50-106A, Bethyl Laboratories), claudin-3 (#34-1700, Thermo Fisher Scientific), claudin-4 (#PA5-32354, Thermo Fisher Scientific), or mouse monoclonal antibodies against BCL6 (#sc7388, Santa Cruz Biotechnology), CD3 (#NBP2-53386, Novus Biologicals), CD20 (#NBP2-45454, Novus Biologicals), CD40 (#sc-13128, Santa Cruz Biotechnology), occludin (#sc-133256, Santa Cruz Biotechnology), or a rat monoclonal antibody against MECA79 (#sc-19602, Santa Cruz Biotechnology) diluted in PBS-T containing 2.5% bovine serum albumin.

Techniques: Staining, Immunohistochemistry

Journal: Frontiers in immunology

Article Title: Investigating mammary glands of lactating goats for the presence of tertiary lymphoid organs.

doi: 10.3389/fimmu.2022.941333

Figure Lengend Snippet: FIGURE 3 Representative images of immunohistochemistry against CD20 (B cells) and CD3 (T cells) in large aggregations of lymphocytes in goat mammary glands. Images of the same position are shown. Scale bar, 100 mm.

Article Snippet: The sections were then incubated overnight at 4°C with either rabbit polyclonal antibodies against goat-IgA (#A50-106A, Bethyl Laboratories), claudin-3 (#34-1700, Thermo Fisher Scientific), claudin-4 (#PA5-32354, Thermo Fisher Scientific), or mouse monoclonal antibodies against BCL6 (#sc7388, Santa Cruz Biotechnology), CD3 (#NBP2-53386, Novus Biologicals), CD20 (#NBP2-45454, Novus Biologicals), CD40 (#sc-13128, Santa Cruz Biotechnology), occludin (#sc-133256, Santa Cruz Biotechnology), or a rat monoclonal antibody against MECA79 (#sc-19602, Santa Cruz Biotechnology) diluted in PBS-T containing 2.5% bovine serum albumin.

Techniques: Immunohistochemistry