facs buffer  (R&D Systems)

Journal: bioRxiv

Article Title: Focused Ultrasound Thermal Ablation and CD40 Agonism Reprograms Breast Tumor Immunity to Drive Regression and Memory

doi: 10.64898/2026.03.02.708396

Figure Lengend Snippet: (A) Experimental timeline of T-FUS+αCD40 dosing and harvest for flow cytometry analysis. (B) Average tumor outgrowth of E0771 tumor-bearing mice. Significance assessed by 2-way ANOVA, followed by Holm-Sidak multiple comparison correction. E0771: p<0.05 vs all other groups (specifically, IgG (n=5) vs T-FUS+αCD40 (n=12): p = <0.0001; T-FUS (n=7) vs T-FUS+αCD40: p =0.0843; αCD40 (n=7) vs T-FUS+αCD40: p = 0.0517). (C) Focused view of tumor outgrowth at selected timepoints (day 42-53) highlighting differences between T-FUS+αCD40 non-CRs and CRs, versus αCD40 monotherapy. (D) Waterfall plot depicting CRs in the combination treatment group. (E-H) Absolute number per gram tumor and percentage of CD8 + and CD4 + T cells in the tumor and (I-L) spleen. M) tSNE plot on CD45 + cells in the TDLN. (N-Q) Absolute number and percentage of CD8 + and CD4 + T cells in the TDLN. (R) Ratio between CD8 + and CD4 + T cells in the TDLN. Significance assessed using a Welch ANOVA or Welch’s T test. *p < 0.05, **p < 0.01 vs. T-FUS+αCD40 CRs.

Article Snippet: Following, cells were fixed with anti-mouse CD16/32 (ThermoFisher #14-0161-86) to block Fc gamma receptors for 15 mins at 4oC, then centrifuged and washed twice with 1X Flow Cytometry Staining Buffer (FACS; R&D Systems #FC001).

Techniques: Flow Cytometry, Comparison

Journal: International Journal of Molecular Sciences

Article Title: The Function and Role of Intercellular Adhesion Molecule 2 in Dental Pulp Cells and Tissue

doi: 10.3390/ijms262412006

Figure Lengend Snippet: ICAM2 localization in rat dental pulp tissue and ICAM2 expression in HDPCs. ( A ) The mRNA expression of ICAM1 , ICAM2 (black column), ICAM3 , ICAM4 , ICAM5 in HDPC-5Y was assessed by quantitative RT-PCR (qRT-PCR). It was normalized against β-actin expression (means ± SD; n = 4; ** p < 0.01). ( B ) Hematoxylin-eosin (H&E) staining of tissue sections (sagittal sections) of mandibular first molars from Wistar rats. The right panel is the higher magnification view of boxed area in the left panel. PU: dental pulp tissue; De: Dentin. Bars, 100 μm. ( C – E ) Immunofluorescence (IF) staining of ICAM2 in the normal dental pulp tissue ( C ). The higher magnification view of boxed area in ( C , D ). Positive staining was indicated by arrow heads (odontoblasts) and arrow (dental pulp cells). Negative control: rabbit IgG (cIgG; ( E )). Nuclei were stained with DAPI (Blue). Bars, 100 μm. ( F , G ) The expression of ICAM2 in HDPC-5Y was examined by IF staining. Anti-ICAM2: Green ( F ), anti-rabbit IgG (control IgG: cIgG; ( G )). Nuclei were stained with DAPI (Blue). Bars, 100 μm. Arrow heads indicate ICAM2-positive HDPCs ( F ). ( H ) The gene expression of ICAM2 in three HDPCs (HDPC-5Y, 5L, and 5I) was examined by semi-qRT-PCR. It was normalized against GAPDH expression. ( I ) The expression intensities of ICAM2 (CD102) of HDPC-5Y (orange line) were demonstrated by flow cytometry. In the gated region, positive cells. Gray line indicates negative control (rabbit IgG).

Article Snippet: Cells (2 × 10 5 /tube) were prepared as a single cell suspension by trypsin/EDTA digestion and resuspended in flow cytometry buffer (R&D Systems, Minneapolis, MN, USA), then incubated with antibodies (10 mg/mL) specific for surface markers or isotype control antibodies (10 mg/mL) on ice for 45 min. Anti-CD102 (ICAM2)-PE antibodies (eBioscience, San Diego, CA, USA) and mouse IgG-PE isotype control were used.

Techniques: Expressing, Quantitative RT-PCR, Staining, Immunofluorescence, Negative Control, Control, Gene Expression, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: The Function and Role of Intercellular Adhesion Molecule 2 in Dental Pulp Cells and Tissue

doi: 10.3390/ijms262412006

Figure Lengend Snippet: ICAM2 localization in rat dental pulp tissue and ICAM2 expression in HDPCs. ( A ) The mRNA expression of ICAM1 , ICAM2 (black column), ICAM3 , ICAM4 , ICAM5 in HDPC-5Y was assessed by quantitative RT-PCR (qRT-PCR). It was normalized against β-actin expression (means ± SD; n = 4; ** p < 0.01). ( B ) Hematoxylin-eosin (H&E) staining of tissue sections (sagittal sections) of mandibular first molars from Wistar rats. The right panel is the higher magnification view of boxed area in the left panel. PU: dental pulp tissue; De: Dentin. Bars, 100 μm. ( C – E ) Immunofluorescence (IF) staining of ICAM2 in the normal dental pulp tissue ( C ). The higher magnification view of boxed area in ( C , D ). Positive staining was indicated by arrow heads (odontoblasts) and arrow (dental pulp cells). Negative control: rabbit IgG (cIgG; ( E )). Nuclei were stained with DAPI (Blue). Bars, 100 μm. ( F , G ) The expression of ICAM2 in HDPC-5Y was examined by IF staining. Anti-ICAM2: Green ( F ), anti-rabbit IgG (control IgG: cIgG; ( G )). Nuclei were stained with DAPI (Blue). Bars, 100 μm. Arrow heads indicate ICAM2-positive HDPCs ( F ). ( H ) The gene expression of ICAM2 in three HDPCs (HDPC-5Y, 5L, and 5I) was examined by semi-qRT-PCR. It was normalized against GAPDH expression. ( I ) The expression intensities of ICAM2 (CD102) of HDPC-5Y (orange line) were demonstrated by flow cytometry. In the gated region, positive cells. Gray line indicates negative control (rabbit IgG).

Article Snippet: Briefly, 6 × 10 6 HDPCs were suspended in Flow Cytometry Staining Buffer (R&D Systems, Minneapolis, MN, USA) and centrifuged at 300× g for 10 min, resuspended in 80 μL of MACS buffer (Miltenyi Biotec) with 20 μL CD102 microbeads, and incubated in the dark at 4 °C for 15 min.

Techniques: Expressing, Quantitative RT-PCR, Staining, Immunofluorescence, Negative Control, Control, Gene Expression, Flow Cytometry