dapi fluoromount-g clear mounting media  (Thermo Fisher)

Journal: bioRxiv

Article Title: Tissue-specific tolerance mechanisms and lymph node co-drainage converge to shape T cell immunity in the upper digestive system and regulate pancreatic cancer progression

doi: 10.64898/2025.12.02.691870

Figure Lengend Snippet: A-B. Expression levels of OVA in the liver ( A ) and pancreas ( B ) of mice with indicated genotypes ( n = 3-10 per group, as indicated by symbols). C-D . Hepatic ( C ) and pancreatic ( D ) tissue lysates from mice with indicated OVA form and their CRE-negative controls were immunoblotted using antibodies against FLAG, and β-ACTIN as a loading control. E . Sections of pancreas (top) and liver (bottom) from mice with indicated genotypes stained for FLAG and DAPI (nuclei). Scale bar represents 20μm. F-H. OVA expression levels in the liver, pancreas and duodenum tissue of mice with indicated genotypes, showing tissue-specific expression restricted to the intended sites ( n = 3-10 per group, as indicated by symbols). I. Division index of OT-II cells in the indicated LNs 6 days after transfer into Ins1 sOVA , Ins1 cOVA and Ins1 tmOVA mice ( n = 4-7 per group, as indicated by symbols). J . Hepatic tissue lysates from Alb sOVA mice treated with or without tamoxifen and B6 mice were immunoblotted using antibodies against FLAG and β-ACTIN. K. OVA expression levels in liver tissue of Alb sOVA mice treated with or without tamoxifen and sOVA mice infected with adeno-Cre virus and pancreas of Ptf1a sOVA mice ( n = 3-5 per group, as indicated by symbols). L . Hepatic tissue lysates from Alb sOVA mice without tamoxifen treatment and pancreatic lysates from tamoxifen-treated Ptf1a sOVA mice were immunoblotted using antibodies against FLAG and β-ACTIN. Data are pooled from two independent experiments in A, B, F-I. * p<0.05, ** p<0.01, *** p<0.001 by t-test.

Article Snippet: The following day, slides were washed three times with PBST (PBS with 0.05% Tween-20) for 15 min each, incubated with appropriate secondary antibodies for 1 h at RT, washed again three times in PBST, and mounted with DAPI Fluoromount G Clear Mounting Media (SouthernBiotech).

Techniques: Expressing, Control, Staining, Infection, Virus

Journal: bioRxiv

Article Title: Tissue-specific tolerance mechanisms and lymph node co-drainage converge to shape T cell immunity in the upper digestive system and regulate pancreatic cancer progression

doi: 10.64898/2025.12.02.691870

Figure Lengend Snippet: A-D. Frequencies of OT-I cells among total CD8 + cells ( A ), OT-II cells among total CD4 + cells ( B ), GzmB + among total OT-I cells ( C ) and T-BET + among total OT-II cells ( D ) in the pancreas of orally T1L-infected or vehicle-infected Ptf1a sOVA mice 7 days post transfer ( n = 6-8 per group, as indicated by symbols). E. Frozen sections of the pancreas from T T1L-infected or vehicle-infected Ptf1a sOVA mice 14 days post transfer stained for CD3, CD8, CD45.1 and DAPI (nuclei). Scale bar represents 100 μm. F-I. Sections of the pancreas from T1L-or mock-infected Ptf1a sOVA mice 14 days post transfer stained with H&E ( F ) or stained for apoptotic cells and DAPI ( H ). Scale bars represent 100 μm and 200 μm, respectively. Quantification of histological score ( G ) and apoptotic index ( I ) ( n = 5-8 per group, as indicated by symbols). Data are pooled from two independent experiments in A-D, G, I. * p<0.05, ** p<0.01, *** p<0.001 by t-test.

Article Snippet: The following day, slides were washed three times with PBST (PBS with 0.05% Tween-20) for 15 min each, incubated with appropriate secondary antibodies for 1 h at RT, washed again three times in PBST, and mounted with DAPI Fluoromount G Clear Mounting Media (SouthernBiotech).

Techniques: Infection, Staining

Journal: bioRxiv

Article Title: Tissue-specific tolerance mechanisms and lymph node co-drainage converge to shape T cell immunity in the upper digestive system and regulate pancreatic cancer progression

doi: 10.64898/2025.12.02.691870

Figure Lengend Snippet: A, D . Schematic of orthotopic PDAC model for assessing T cell fate in the LNs ( A ) and tumor progression ( D ). B-C . Frequencies of GzmB + among total OT-I cells ( B ) or T-BET + among total OT-II ( C ) cells in indicated LNs of mice receiving sOVA- or cOVA-expressing tumor cells ( n = 4-5 per group, as indicated by symbols). E. Representative photographs of tumors removed from mice with indicated tumor implants and T1L infection status. F-G. Weights ( F ) and sizes ( F ) of tumors from mice in ( E ) ( n = 4-10 per group, as indicated by symbols). H. Tumor frozen sections from mice in ( E ) were stained for CD3, CD8, CD45.1 and DAPI (nuclei). Scale bar represents 50 μm. I-J. Weights ( I ) and sizes ( J ) of tumors from mice receiving sOVA-expressing KPC cells intragastrically or intranasally infected with T1L ( n = 5 per group). Data are pooled from two independent experiments in F, G. Data represent one experiment in B, C, I, J. * p<0.05, ** p<0.01, *** p<0.001 by t-test.

Article Snippet: The following day, slides were washed three times with PBST (PBS with 0.05% Tween-20) for 15 min each, incubated with appropriate secondary antibodies for 1 h at RT, washed again three times in PBST, and mounted with DAPI Fluoromount G Clear Mounting Media (SouthernBiotech).

Techniques: Expressing, Infection, Staining

Journal: bioRxiv

Article Title: Tissue-specific tolerance mechanisms and lymph node co-drainage converge to shape T cell immunity in the upper digestive system and regulate pancreatic cancer progression

doi: 10.64898/2025.12.02.691870

Figure Lengend Snippet: A. Plots of OT-II cells in indicated LNs of orally T1L-infected or vehicle-infected Alb tmOVA mice, representative of gating applies in ( B ) and ( C ). B-C. Frequencies of FOXP3 + ( B ) or T-BET + ( C ) among total OT-II cells in indicated LNs 72 h after transfer into mice in ( A ) ( n = 5-6 per group, as indicated by symbols). D. Plots of OT-I cells in indicated LNs of T1L-or vehicle-infected Alb tmOVA mice, representative of gating applied in ( E ) – ( G ). E-G. Frequencies of total GzmB + ( E ), IFNψ + GzmB + ( F ) or IFNψ + TNFα + ( G ) among total OT-I cells in indicated LNs 72 h after transferred into mice in ( D ) ( n = 4 per group). H-K . Frequencies of OT-I cells among total CD8 + cells ( H ), OT-II cells among total CD4 + cells ( I ), GzmB + among total OT-I cells ( J ) and T-BET + among total OT-II cells ( K ) in the liver of T1L- or mock-infected Alb tmOVA mice 7 days post transfer ( n = 5 per group). L. Sections of the liver from orally T1L-infected or vehicle-infected Alb tmOVA mice 14 days post transfer stain with H&E. Scale bar represents 400 μm. M. Frozen sections of the liver from T1L-infected or vehicle-infected Ptf1a sOVA mice stained for CD3, CD8, CD45.1 and DAPI (nuclei). Scale bar represents 200 μm. N. Sections of the liver from T1L-infected or vehicle-infected Alb tmOVA mice 14 days post infection stained for apoptotic cells. Scale bar represents 100 μm. Data are representative of two independent experiments in B, C, E-K. * p<0.05, ** p<0.01, *** p<0.001 by t-test.

Article Snippet: The following day, slides were washed three times with PBST (PBS with 0.05% Tween-20) for 15 min each, incubated with appropriate secondary antibodies for 1 h at RT, washed again three times in PBST, and mounted with DAPI Fluoromount G Clear Mounting Media (SouthernBiotech).

Techniques: Infection, Staining