Journal: Nature Communications

Article Title: FAM3D is essential for colon homeostasis and host defense against inflammation associated carcinogenesis

doi: 10.1038/s41467-020-19691-z

Figure Lengend Snippet: a Heatmaps of differentially expressed genes enriched in host defense responses. b mRNA expression levels of antibacterial peptides in the colons of WT or Fam3D −/− mice measured by real-time PCR. n = 8. c Reg3γ protein levels measured by immunofluorescent staining. n = 4 for each group. d GSEA analysis on Fam3D −/− vs WT EBSeq log 2 FC expression data of colonic epithelial samples. e Immunofluorescent staining for Fam3D and MUC2 in normal colon tissue. Data is representative of one experiment repeated three independent times. f Double staining of Acian blue and PAS. Yellow arrow: Acian blue positive materials; orange arrow: PAS single positive cells; orange star: dilated goblet cells, goblet cells enumerated in each crypt (10–20 crypts from each colon) and pixels of Alcian blue staining areas. Alcian blue staining areas and fluorescence intensity quantified by Image J based on six distal colon sections of four independent mice from each group. g Staining of high iron diamine/Alcian blue, dark brown, and black were sulfomucins, whereas blue staining indicates sialomucins, sulfomucin positive cells enumerated in each crypt (10–20 crypts from each colon) and the ratio of sialomucins to sulfomucins is calculated. Data is presented as the mean ± SEM. Scale bar = 50 μm. h Representative Alcian blue staining of Carnoy’s-fixed colonic sections. The thickness of the inner mucus layer (light blue band between luminal content and the mucosa) was quantified (5–10 fields from each colon) by Image J. n = 8. i Representative dual staining for UEA-I (immunofluorescence, green) and bacteria (fluorescence in situ hybridization, red), the latter using the universal EUB338 probe, on Carnoy’s-fixed colonic sections. n = 8. j PAS/Alcian blue staining of the colons from WT and Fam3D −/− mice with different ages. n = 4. Data is representative of one experiment repeated three independent times. k Quantitation of crypt length of WT and Fam3D −/− mice at different ages. Data is presented as the mean ± SEM. Statistical significance was determined by unpaired, two-tailed Student’s t test.

Article Snippet: Rat anti-EpCAM antibody (sc-53532), mouse anti-β-catenin antibody (sc-7963), rabbit anti-mouse Mucin 2 (MUC2) antibody (H300) were purchased from Santa Cruz (Dallas, TX).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Staining, Double Staining, Fluorescence, Immunofluorescence, Bacteria, In Situ Hybridization, Quantitation Assay, Two Tailed Test

Journal: Mucosal immunology

Article Title: Hypoxia-inducible factor 1 in dendritic cells is crucial for the activation of protective regulatory T cells in murine colitis.

doi: 10.1038/mi.2015.67

Figure Lengend Snippet: Figure 6 Dendritic cell-specific knockout of hypoxia-inducible factor-1a (HIF-1a) stimulates mucosal epithelium to increased production of mucins in dextran sodium sulfate (DSS) colitis. Colonic mRNA expression of (a) MUC1, (b) MUC2, and (c) MUC3 was significantly higher in DSS-treated CD11cCre/HIF-1a þ f/ þ f mice than in HIF-1a þ f/ þ f DSS-treated mice or in mice not treated with DSS. (d) Sections (4 mm) of colon tissue were stained with MUC2 (red) and 4’,6-diamidino-2-phenylindole (DAPI; blue). After treatment with DSS, the amount of colonic MUC2 protein was higher in CD11cCre/HIF- 1a þ f/ þ f mice than in HIF-1a þ f/ þ f mice (original magnification 200; scale bar ¼ 50 mm). N ¼ 5–7 per group. *Po0.05; **Po0.01; ****Po0.0001. C, control; NS, not significant.

Article Snippet: MUC2 was detected with a rabbit anti-mouse MUC2 antibody (Santa Cruz Biotechnology).

Techniques: Knock-Out, Expressing, Staining, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Intestinal Dysbiosis Amplifies Acetaminophen-Induced Acute Liver Injury

doi: 10.1016/j.jcmgh.2020.11.002

Figure Lengend Snippet: Intestinal barrier impairment on APAP challenge. ( A ) Representative immunofluorescence of colon of WT and Nlrp6 -/- mice staining against zonula occludens-1. ( B ) Western blot analyses and ( C ) quantification of occludin protein expression in ileum of representative WT and Nlrp6 -/- mice after 12 hours of NaCl (n = 3) or APAP (n = 5) treatment. ( D ) Colonic mucus layers and bacterial colonization shown by immunofluorescent staining against Muc2 and fluorescence in situ hybridization using EuBac- probe for eubacteria in colon of WT and Nlrp6 -/- mice. ( E ) Enzyme-linked immunosorbent assay of fecal albumin concentrations in freshly collected fecal pellets from APAP-treated WT (n = 9) and Nlrp6 -/- (n = 7) mice and controls (WT: n = 4, Nlrp6 -/- : n = 3). ( F ) Quantitative analysis of thickness of colonic mucus layers in immunofluorescent stainings (NaCl: WT: n = 4, Nlrp6 -/- : n = 2; APAP: WT: n = 3, Nlrp6 -/- : n = 5) 24 hours after APAP administration. All data are presented as mean ± standard error of mean and considered significant at ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001, respectively (unpaired Student t test).

Article Snippet: After paraffin embedding, mucus and gut bacteria were stained using an anti-Muc2 primary antibody (Santa Cruz Biotechnology, Dallas, TX; sc-15334) and a fluorescence in situ hybridization probe against eubacteria (16S rRNA: 5'-GCTGCCTCCCGTAGGAGT-3').

Techniques: Immunofluorescence, Staining, Western Blot, Expressing, Fluorescence, In Situ Hybridization, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Microbiology

Article Title: Introduction of Colonic and Fecal Microbiota From an Adult Pig Differently Affects the Growth, Gut Health, Intestinal Microbiota and Blood Metabolome of Newborn Piglets

doi: 10.3389/fmicb.2021.623673

Figure Lengend Snippet: CMT improved development and immune function of the gut in piglets. (A) Hematoxylin-eosin staining images (40 × magnification) and scanning electron microscope images (300 × magnification) of ileal sections from piglets on day 28. (B) Development index of the ileum (height of ileal villi, depth of ileal crypts, and muscular thickness). (C) Protein levels of tight junction proteins in the ilea of piglets. The expression levels of proteins were analyzed by western blotting. (D) Protein levels of toll-like receptor 2 and 4 (TLR2 and TLR4) in the ilea of piglets. (E) Diamine oxidase content in the blood of piglets. (F) Mucosal thickness in the colons of piglets. (G) Immunohistochemical staining of the Mucin 2 (MUC2) protein in colonic sections. (H) mRNA level of MUC2. The expression level was detected by qRT-PCR ( n = 6). The data are presented as the mean ± S.D. ∗∗∗ Indicates P < 0.001; ∗∗ indicates P < 0.01, and ∗ indicates P < 0.05 compared with the control; ### indicates P < 0.001, ## indicates P < 0.01, and # indicates P < 0.05 between the FMT and CMT groups.

Article Snippet: A rabbit polyclonal anti-MUC2 antibody was purchased from Proteintech (#27675-1-AP).

Techniques: Staining, Microscopy, Expressing, Western Blot, Immunohistochemical staining, Quantitative RT-PCR, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Congruence of Transcription Programs in Adult Stem Cell-Derived Jejunum Organoids and Original Tissue During Long-Term Culture

doi: 10.3389/fcell.2020.00375

Figure Lengend Snippet: Organoids derived from adult intestinal stem cells show intrinsic programming to differentiate into different epithelial cell lineages and express small intestine-specific genes. (A) Cell type-specific transcripts for Crypt Base Columnar (CBC) and Stem cells, Label-retaining (LRC) +4 cells, Proliferation (Prol), Niche factors (NF), Paneth cells (PC), Goblet cells (GC), Enteroendocrine cells (EEC), and Absorptive cells or enterocytes (Abs/EC). (B) Overlaying the NCBI gene tracks of MUC2 NC_010444.4 on chromosome 2 at location 689363–719542bp (green area), shows identical overlap with the mapped reads and coverage (Cov) from organoid and tissue samples, but not in IPEC-J2 (ENS; Ensembl reference genome). To confirm MUC2 protein translation and subsequent mucus formation, Carnoy fixed tissue, organoid, and IPEC-J2 samples were stained with PAS/Alcian blue (left) and porcine anti-MUC2 (right; black and white size bars indicate 100 μm). (C) Most small intestine-specific genes identified in the human protein atlas are also expressed in porcine jejunum tissue and their derived organoids (>74%), whereas fewer are expressed in ileum organoids (52%). IPEC-J2 only expressed 32% of the small intestine-specific genes. (D) Organoid transcriptomes from jejunum and ileum were compared to identify differences in expressed genes, showing large overlap of genes (Venn), but also some differences. The different genes were analyzed using TOPPfun to identify putative differences in gene ontology and pathways (TMT, Transmembrane transport; PSP, Peptidyl-serine phosphorylation).

Article Snippet: After cutting 5 μM-thick sections and subsequent drying on glass slides, sections were rehydrated, blocked in 5% normal goat serum, and stained using anti-MUC2 antibody (1:200, AB_1950958, GeneTex) overnight.

Techniques: Derivative Assay, Staining, Phospho-proteomics

Journal: Molecular Medicine

Article Title: Evaluation of two laboratory model methods for diarrheal irritable bowel syndrome

doi: 10.1186/s10020-022-00599-x

Figure Lengend Snippet: The primers used in this experiment

Article Snippet: Antibodies zonulae occluden-1 (ZO-1) and mucin 2 (Muc2) were purchased from Boster Bioengineering Co., LTD. (PB9234, BM5029).

Techniques: Sequencing

Journal: Molecular Medicine

Article Title: Evaluation of two laboratory model methods for diarrheal irritable bowel syndrome

doi: 10.1186/s10020-022-00599-x

Figure Lengend Snippet: Intensity of ZO-1 and MUC2 (green) in ileal and colonic tissues of rats in each group. Nuclei stained with DAPI (blue) (200×) ( A ), mRNA expression levels of ZO-1 ( B ), MUC2 ( C ), OCLN ( D ), CLDN4 ( E ) in each group of rats.* p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Antibodies zonulae occluden-1 (ZO-1) and mucin 2 (Muc2) were purchased from Boster Bioengineering Co., LTD. (PB9234, BM5029).

Techniques: Staining, Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Sialic acid plays a pivotal role in licensing Citrobacter rodentium's transition from the intestinal lumen to a mucosal adherent niche.

doi: 10.1073/pnas.2301115120

Figure Lengend Snippet: Fig. 1. Citrobacter rodentium resides within the colonic mucus and catabolizes sialic acid. (A) C. rodentium localizes to the mucus layer. Representative immunofluorescence staining of mouse colonic tissue infected with C. rodentium. A colon cross-section (green panel) was stained with DAPI to detect DNA (blue), anti-C. rodentium (red) to visualize C. rodentium, and anti-Muc2 to visualize mucus (green). The gray panel is the enlarged view of the boxed region within the cross-section, original magnification = 200×. The orange panel is a magnified image indicating a subpopulation of C. rodentium localized to the inner mucus and traversing the mucus (arrowheads), with a separate image showing C. rodentium staining independently (red channel), original magnification = 630×. (Scale bar, 15 μm.) (B) C. rodentium uses sialic acid as a sole carbon source for growth. C. rodentium growth was measured by optical density (OD600) at 20-min intervals over 24 h at 37 °C in M9 minimal medium supplemented with 0.2% N-acetylneuraminic acid (sialic acid) or purified mucins. Data are presented as averages of cell growth (n = 9) from three independent experiments.

Article Snippet: For visualizing C. rodentium localization in the mucus, methacarn- fixed mouse distal colons were stained with the following primary antibodies—rat anti- C. rodentium Tir (gift from W. Deng), rabbit anti- Muc2 (Boster), and rabbit anti- Muc2 (Novus), which were then probed with Alexa Fluor 488- conjugated donkey anti- rabbit IgG (Life Technologies) and Alexa Fluor 568–conjugated donkey anti- rat IgG (Life Technologies).

Techniques: Immunofluorescence, Staining, Infection, Purification

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Sialic acid plays a pivotal role in licensing Citrobacter rodentium's transition from the intestinal lumen to a mucosal adherent niche.

doi: 10.1073/pnas.2301115120

Figure Lengend Snippet: Fig. 2. Sialic acid in the colon is mainly derived from mucus produced by goblet cells and widely expressed before and during C. rodentium infection. (A) Representative immunofluorescence staining of sialic acid on murine colonic sections with and without C. rodentium infection. Sections were stained with DAPI to detect DNA (blue) and SNA lectin (α2,6-sialic acid binding) to visualize sialic acid. Dotted lines indicate the apical side of the epithelium. Original magnification = 200×. (Scale bar, 50 µm.) (B) Degree of sialylation on Muc2 O-glycans of colonic mucus with (n = 4) and without (n = 4) infection with C. rodentium for 6 d. Released O-glycans from distal intestine were analyzed on PGC-LC-MS/MS. (C) Levels of free sialic acid in fecal contents of mice without (n = 6) and with (n = 6) C. rodentium infection for 6 d. All data are shown as mean ± SEM. Statistical significance calculated by the Mann–Whitney U test (B and C).

Article Snippet: For visualizing C. rodentium localization in the mucus, methacarn- fixed mouse distal colons were stained with the following primary antibodies—rat anti- C. rodentium Tir (gift from W. Deng), rabbit anti- Muc2 (Boster), and rabbit anti- Muc2 (Novus), which were then probed with Alexa Fluor 488- conjugated donkey anti- rabbit IgG (Life Technologies) and Alexa Fluor 568–conjugated donkey anti- rat IgG (Life Technologies).

Techniques: Derivative Assay, Produced, Infection, Immunofluorescence, Staining, Binding Assay, Liquid Chromatography with Mass Spectroscopy, MANN-WHITNEY

Journal: Gut Microbes

Article Title: Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus

doi: 10.1080/19490976.2025.2494717

Figure Lengend Snippet: The ALI model produces a physiologically relevant mucus layer. a, The experimental procedure to generate ALI-derived mucus, created with Biorender.com. b, The formation of the mucus layer after growing ALI cultures for 7 days, 14 days and 21 days. Arrows indicate the mucus layer. The average mucus thickness at 14 days and 21 days is 114 ± 28 µm, and 140 ± 25 µm, respectively. c, Immunostaining of Muc2 -/- ALI (at 21 days). ALI cross-sections were stained with DAPI (blue), anti-Muc2 (green) and anti-E-cadherin (white). d, Proteomics analysis of gel-forming mucins present in mouse ALI-derived mucus. e-f, Pie chart of acidic (e) and various subsets (f) of mucin-type O-glycans in colonic ALI-derived mucus. g, MALII (red) staining of Muc2 +/+ ALI culture. Scale bar, 200 µm.

Article Snippet: Muc2 degradation products were detected by immunoblotting using an anti-Muc2 rabbit polyclonal antibody (Novus Biologicals, NBP1–31231) (1:1,000) which targets an epitope on the C terminus of mucin 2 from both human and mouse origin.

Techniques: Derivative Assay, Immunostaining, Staining

Journal: Gut Microbes

Article Title: Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus

doi: 10.1080/19490976.2025.2494717

Figure Lengend Snippet: The ALI model is able to recapitulate pathogen-mucus interaction in vivo . a, C. rodentium infection of Muc2 +/+ ALI cultures for 6 h and 10 h. Scale bar, 200 µm. b, C. rodentium infection of Muc2 -/- ALI culture for 6 h. ALI cross-sections were stained with anti- C. rodentium LPS (red), Ulex europaeus agglutinin-1 (UEA-1, green), DAPI (blue) and E-cadherin (white). Scale bar, 200 µm. White arrows indicate C. rodentium present in the mucus. Yellow arrows indicate C. rodentium close to IECs. Yellow arrowheads indicate cell sloughing.

Article Snippet: Muc2 degradation products were detected by immunoblotting using an anti-Muc2 rabbit polyclonal antibody (Novus Biologicals, NBP1–31231) (1:1,000) which targets an epitope on the C terminus of mucin 2 from both human and mouse origin.

Techniques: In Vivo, Infection, Staining

Journal: Gut Microbes

Article Title: Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus

doi: 10.1080/19490976.2025.2494717

Figure Lengend Snippet: Sialic acid enhances C. rodentium ’s ability to degrade ALI-derived mucus and infect IECs. a, Diagram of mucus degradation assay in vitro , created with Biorender.com. b, Mucus degradation assay using the supernatant from C. rodentium grown in media containing 0.45% glucose (Glc) or sialic acid (SA). Mucus incubated with C. rodentium supernatants was loaded onto 3–8% Tris-acetate gels and run through electrophoresis. Proteins were visualized by western blot using an anti-Muc2 antibody. c, Quantification analysis of degraded Muc2 band to total Muc2 band, ***, p < 0.001; ****, p < 0.0001. d, Immunostaining of ALI cultures infected with C. rodentium in the presence of glucose or sialic acid. ALI cross-sections were stained with anti- C. rodentium LPS (red), UEA-1(green), DAPI (blue) and E-cadherin (white). Yellow arrows indicate C. rodentium close to IECs. Yellow arrowheads indicate cell sloughing. Scale bar, 200 µm.

Article Snippet: Muc2 degradation products were detected by immunoblotting using an anti-Muc2 rabbit polyclonal antibody (Novus Biologicals, NBP1–31231) (1:1,000) which targets an epitope on the C terminus of mucin 2 from both human and mouse origin.

Techniques: Derivative Assay, Degradation Assay, In Vitro, Incubation, Electrophoresis, Western Blot, Immunostaining, Infection, Staining

Journal: Gut Microbes

Article Title: Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus

doi: 10.1080/19490976.2025.2494717

Figure Lengend Snippet: A comparison of the ability of WT, Δpic, ΔespC and ΔpicΔespC C. rodentium strains to infect mouse ALI cultures and to degrade the ALI-derived mucus. a, The ALI cultures were infected with different C. rodentium strains, including WT, Δpic, ΔespC and ΔpicΔespC double mutant ( ΔΔ ). ALI cross-sections were stained with anti- C. rodentium LPS (red), UEA-1(green), DAPI (blue) and E-cadherin (white). White arrows indicate C. rodentium on the top of the mucus. Scale bar, 200 µm. b, Diagram of mucus degradation assay after C. rodentium infection, created with Biorender.com. ALI cultures were infected with different bacterial strains, followed by the collection of mucus from infected cultures. c, mucus from infected ALI was collected and run on a 3%-8% Tris-acetate gel, followed by western blot using an anti-Muc2 antibody.

Article Snippet: Muc2 degradation products were detected by immunoblotting using an anti-Muc2 rabbit polyclonal antibody (Novus Biologicals, NBP1–31231) (1:1,000) which targets an epitope on the C terminus of mucin 2 from both human and mouse origin.

Techniques: Comparison, Derivative Assay, Infection, Mutagenesis, Staining, Degradation Assay, Western Blot

Journal: Gut Microbes

Article Title: Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus

doi: 10.1080/19490976.2025.2494717

Figure Lengend Snippet: Mucus degradation assay using protein secreted by SPATEs expressing E. coli DH5α. a, The degradation of mouse ALI-derived mucus by supernatant from E. coli DH5α expressing CrespC or Crpic . EV, empty vector; L, protein ladder; E, empty lane. Note, the anti-Muc2 antibody shows cross-activity against Pic (~110 kDa). *, p < 0.05. b, The degradation of mouse ALI-derived mucus by supernatant from E. coli DH5α expressing CrespC or CrespC-S251I . c, The degradation of mouse ALI-derived mucus by supernatant from E. coli DH5α expressing CrespC or EPEC espC . Samples from the mucus degradation assay were reduced and run on a 3%-8% Tris-acetate gel, followed by western blot using an anti-C-terminus Muc2 antibody.

Article Snippet: Muc2 degradation products were detected by immunoblotting using an anti-Muc2 rabbit polyclonal antibody (Novus Biologicals, NBP1–31231) (1:1,000) which targets an epitope on the C terminus of mucin 2 from both human and mouse origin.

Techniques: Degradation Assay, Expressing, Derivative Assay, Plasmid Preparation, Activity Assay, Western Blot

Journal: Gut Microbes

Article Title: Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus

doi: 10.1080/19490976.2025.2494717

Figure Lengend Snippet: Recombinant CrEspC is able to degrade mouse ALI-derived mucus. a, The sequence information used to express recombinant CrEspC by E. coli BL21 Star (DE3) system, created with Biorender.com. SS, signal sequence. b, The degradation of mouse ALI-derived mucus by 0.05 µg purified CrEspC for 2 h, 8 h and 24 h. c, The degradation of mouse ALI-derived mucus by different amounts (0.0025, 0.0125, 0.05 µg) of purified CrEspC for 2 h. Samples were reduced and run on a 3%-8% Tris-acetate gel, followed by western blot using an anti-C-terminus Muc2 antibody.

Article Snippet: Muc2 degradation products were detected by immunoblotting using an anti-Muc2 rabbit polyclonal antibody (Novus Biologicals, NBP1–31231) (1:1,000) which targets an epitope on the C terminus of mucin 2 from both human and mouse origin.

Techniques: Recombinant, Derivative Assay, Sequencing, Purification, Western Blot

Journal: BMC Biology

Article Title: Ileum tissue single-cell mRNA sequencing elucidates the cellular architecture of pathophysiological changes associated with weaning in piglets

doi: 10.1186/s12915-022-01321-3

Figure Lengend Snippet: Heterogeneity of epithelial secretory cell subtypes in ileum tissues of piglets. a t-SNE plot displaying 832 epithelial secretory cells (ESCs) separated into 6 subtypes. EC, enterocyte; GC, goblet cell; EEC, enteroendocrine cell; B/OC, BEST4 / OTOP2 cell. b Violin plots showing canonical marker genes across ESC subtypes. See Additional file : Table S2 for all marker genes. c Heatmap reflecting differentially expressed transcription factors and cell-type-specific genes in ESC subtypes. d Immunofluorescence (IF) staining assay of GC marker MUC2 (green) and EEC markers CCK (red) and GCG (red) ( n =6); arrow, target cell type; scale bars, 50 μm. e Differentiation pseudotime trajectory analysis of ESC subtypes. Predicted secretory lineage cells include GC, EEC, and B/OC; enterocytes (EC) represent absorptive cells. Red arrow indicates the direction of differentiation. f The differentially expressed genes (rows) along the pseudotime (columns) of secretory and absorptive cells clustering hierarchically into four profiles. The representative gene functions and pathways of each profile are shown

Article Snippet: The primary antibodies were involved in IF staining: rabbit anti- MUC2 (Biorbyt Ltd., orb372331, 1:200); rabbit polyclonal cholecystokinin ( CCK ) antibody (Origene, #20078, 1:150); mouse anti- GCG monoclonal antibody (Novus Biologicals, NBP2-21803, 1:2000); rabbit anti -CD45 (intracellular domain) (Cell Signaling, #13917, 1:300); mouse anti-Sybdecan-1 (Abcam, #ab181789, 1:250); rabbit anti-IgA alpha chain (Abcam, #ab97000, 1:400); rabbit anti- CD3 (Abcam, #ab135372, 1:200); rabbit anti- TNF alpha (Abcam, #EPR19147, 1:1,000); rabbit anti -IL1β mAb (Cell Signaling, #D3U3E, 1:500); rabbit anti- BID (Abcam, #ab32060, 1:100); rabbit anti- CD3 -(E4T1B) XP® mAb (CST, #78588, 1:400); mouse anti-CD8α (RPA-T8) mAb (CST, #55397, 1:400); rabbit anti-granzyme B (D2H2F) mAb (CST, #17215, 1:800); rabbit anti-perforin (E9F7N) mAb (Cell Signaling, #44865, 1:100); and rabbit anti- ITGAE/CD103 (EP206) mAb (Cell Signaling, #95835, 1:150).

Techniques: Marker, Immunofluorescence, Staining