Journal: Cancers

Article Title: The Thermal Dose of Photothermal Therapy Generates Differential Immunogenicity in Human Neuroblastoma Cells

doi: 10.3390/cancers14061447

Figure Lengend Snippet: PBNP-PTT generates a thermal dose window of immunogenic cell death in SH-SY5Y and LAN-1 cells in vitro. Three million ( A – C ) SH-SY5Y or ( D – F ) LAN-1 cells were exposed to various thermal doses using PBNP-PTT. After 24 h, cells were analyzed for ( A , D ) intracellular ATP, ( B , E ) HMGB1 release, and ( C , F ) surface calreticulin expression, represented as median fluorescence intensity (MFI). Inset values in the histograms denote the thermal dose. The extent of ICD as measured by its correlates is more pronounced in SH-SY5Y cells compared with LAN-1 cells. Ordinary one-way ANOVA was used to calculate significance between vehicle and different thermal doses and laser alone for HMB1 analysis. n = 2/group; * p < 0.03, ** p < 0.002, *** p < 0.0002, **** p < 0.0001.

Article Snippet: After 24 h incubation at 37 °C post-PBNP-PTT, cells were harvested and stained with Zombie Aqua Fixable viability dye (Biolegend, #423102), blocked with human TruStain Fc block (Biolegend, #422302), and stained with fluorescent antibodies against calreticulin (Abcam, #ab83220), CD80 (Biolegend, #305238), CD86 (Biolegend, #305428), PD-L1 (Biolegend, #329714), B7-H3 (Biolegend, #351010), HLA-ABC (Biolegend, #311432), HLA-DR (Biolegend, #307633), PVR (Biolegend, #337628), and GD2 (Biolegend, #357308).

Techniques: In Vitro, Expressing, Fluorescence

Journal: Cancers

Article Title: The Thermal Dose of Photothermal Therapy Generates Differential Immunogenicity in Human Neuroblastoma Cells

doi: 10.3390/cancers14061447

Figure Lengend Snippet: PBNP-PTT triggers greater immunophenotypic changes in MYCN-non-amplified SH-SY5Y cells than MYCN-amplified LAN-1 neuroblastoma cell line in vitro. SH-SY5Y (blue) and LAN-1 (green) cells were treated with varied thermal doses via PBNP-PTT and analyzed for ( A ) % live cells ( B ) intracellular ATP, ( C ) secreted HMGB1, and cell surface expression levels of ( D ) calreticulin, ( E ) CD80, ( F ) CD86, ( G ) PD-L1, ( H ) B7-H3, ( I ) HLA-ABC, ( J ) HLA-DR, ( K ) PVR, and ( L ) GD2. Data represent mean ± SD ( n = 2 independent samples).

Article Snippet: After 24 h incubation at 37 °C post-PBNP-PTT, cells were harvested and stained with Zombie Aqua Fixable viability dye (Biolegend, #423102), blocked with human TruStain Fc block (Biolegend, #422302), and stained with fluorescent antibodies against calreticulin (Abcam, #ab83220), CD80 (Biolegend, #305238), CD86 (Biolegend, #305428), PD-L1 (Biolegend, #329714), B7-H3 (Biolegend, #351010), HLA-ABC (Biolegend, #311432), HLA-DR (Biolegend, #307633), PVR (Biolegend, #337628), and GD2 (Biolegend, #357308).

Techniques: Amplification, In Vitro, Expressing

Journal: Cell & Bioscience

Article Title: M1-BMDMs with Wnt5a deletion attenuate liver fibrosis by suppression of Wnt5a/Frizzled 2 axis in hepatic progenitors

doi: 10.1186/s13578-025-01467-x

Figure Lengend Snippet: M1-BMDM Wnt5a −KD injection inhibits HPCs activation and DR. ( a ) CK7 and CK19 immunostaining (×200). ( b ) EpCam and Sox9 immunostaining (×200). ( c ) Protein and mRNA expression levels of CK7, CK19, EpCAM and SOX9. ( d ) OV6/CK7 immunofluorescence co-staining (×400), and the costaining cells ratio of OV6/CK7. ( e ) OV6/CK19 immunofluorescence co-staining (×400), and the costaining cells ratio of OV6/CK19. ( f ) EpCam/CK19 immunofluorescence co-staining (×400), and the costaining cells ratio of EpCam/CK19. ( g ) Sox9/CK19 immunofluorescence co-staining (×400), and the costaining cells ratio of Sox9/CK19. * P < 0.05; ** P < 0.01

Article Snippet: The following antibodies were utilized for immunostaining and immunoblot analysis: rabbit polyclonal antibody anti-CD68 (ab125212), mouse monoclonal antibody anti-alpha smooth muscle actin (α-SMA; ab5694/ab124964), rabbit polyclonal antibodies anti-cytokeratin 7 (CK7; ab181598), EpCam (ab71916) and Sox9 (ab185230) (Abcam, Cambridge, UK); mouse monoclonal antibodies anti-transforming growth factor β1 (TGF-β1; sc-130348), OV6 (sc-101863) and Wnt5a (sc-365370) (Santa Cruz Biotechnology, Inc., CA, USA); mouse monoclonal antibody anti-Wnt5a ( SAB53430 ) (Signal way Antibody, Maryland, USA); mouse monoclonal antibody anti-Frizzled 2 (Fzd2; 24272-1-AP), rabbit polyclonal antibodies anti-cytokeratin 19 (CK19; 10712-1-AP) and glyceraldehyde-3-phosphate dehydrogenase antibody (GAPDH) (Proteintech Group Inc., Chicago, IL, USA); IRDye 800CW-conjugated donkey anti-mouse IgG (H + L) and IRDye 680RD-conjugated donkey anti-rabbit IgG (H + L) (LI-COR Bioscience, San Jose, CA, USA); 4’,6-diamidino-2-phenylindole (DAPI; C1002) (Beyotime, Shanghai, China).

Techniques: Injection, Activation Assay, Immunostaining, Expressing, Immunofluorescence, Staining

Journal: Cell & Bioscience

Article Title: M1-BMDMs with Wnt5a deletion attenuate liver fibrosis by suppression of Wnt5a/Frizzled 2 axis in hepatic progenitors

doi: 10.1186/s13578-025-01467-x

Figure Lengend Snippet: M1-BMDM Wnt5a −OE injection promotes HPCs activation and DR. ( a ) CK7 and CK19 immunostaining (×200). ( b ) EpCam and Sox9 immunostaining (×200). ( c ) Protein and mRNA expression levels of CK7, CK19, EpCam and Sox9. ( d ) Immunofluorescence costaining of OV6/CK7 (×400) and the costaining area ratio (%). ( e ) Immunofluorescence costaining of OV6/CK19 (×400) and the costaining area ratio (%). ( f ) Immunofluorescence costaining of EpCam/CK19 (×400) and the costaining area ratio (%). ( g ) Immunofluorescence costaining of Sox9/CK19 (×400) and the costaining area ratio (%). * P < 0.05; ** P < 0.01

Article Snippet: The following antibodies were utilized for immunostaining and immunoblot analysis: rabbit polyclonal antibody anti-CD68 (ab125212), mouse monoclonal antibody anti-alpha smooth muscle actin (α-SMA; ab5694/ab124964), rabbit polyclonal antibodies anti-cytokeratin 7 (CK7; ab181598), EpCam (ab71916) and Sox9 (ab185230) (Abcam, Cambridge, UK); mouse monoclonal antibodies anti-transforming growth factor β1 (TGF-β1; sc-130348), OV6 (sc-101863) and Wnt5a (sc-365370) (Santa Cruz Biotechnology, Inc., CA, USA); mouse monoclonal antibody anti-Wnt5a ( SAB53430 ) (Signal way Antibody, Maryland, USA); mouse monoclonal antibody anti-Frizzled 2 (Fzd2; 24272-1-AP), rabbit polyclonal antibodies anti-cytokeratin 19 (CK19; 10712-1-AP) and glyceraldehyde-3-phosphate dehydrogenase antibody (GAPDH) (Proteintech Group Inc., Chicago, IL, USA); IRDye 800CW-conjugated donkey anti-mouse IgG (H + L) and IRDye 680RD-conjugated donkey anti-rabbit IgG (H + L) (LI-COR Bioscience, San Jose, CA, USA); 4’,6-diamidino-2-phenylindole (DAPI; C1002) (Beyotime, Shanghai, China).

Techniques: Injection, Activation Assay, Immunostaining, Expressing, Immunofluorescence

Journal: Veterinary immunology and immunopathology

Article Title: Flow cytometric analysis of lymphocyte subset kinetics in Bali cattle experimentally infected with Jembrana disease virus.

doi: 10.1016/j.vetimm.2012.06.013

Figure Lengend Snippet: Fig. 1. Representative forward scatter and side scatter gating of PBMC used to identify lymphocytes and their subsets by flow cytometry. Peripheral blood was isolated from cattle on day 0 (pre-infection) and at days 10 (acute phase) and 19 (recovery phase) following experimental JDV infection and PBMC isolated by Ficoll-Paque density separation. Following lymphocyte subset labelling (see following figures C5), cell suspensions were analysed by flow cytometry

Article Snippet: Antibodies and cellular markers Lymphocyte subsets were labelled with 2.5 g/ml ouse anti-bovine CD4 mAb (Serotec, UK), 5 g/ml mouse nti-bovine CD8 mAb (Serotec, UK) or 20 g/ml mouse nti-bovine CD21 mAb (Santa Cruz, USA) as a B-cell marker. n Alexa Fluor 488 (AF488) conjugated goat anti-mouse ross-absorbed secondary antibody (Invitrogen, Australia) as used to detect all reactive mAb antibodies (Table 1).

Techniques: Cytometry, Isolation, Infection

Journal: Veterinary immunology and immunopathology

Article Title: Flow cytometric analysis of lymphocyte subset kinetics in Bali cattle experimentally infected with Jembrana disease virus.

doi: 10.1016/j.vetimm.2012.06.013

Figure Lengend Snippet: Fig. 5. Lymphocyte subset changes related to the febrile response fol- lowing

Article Snippet: Antibodies and cellular markers Lymphocyte subsets were labelled with 2.5 g/ml ouse anti-bovine CD4 mAb (Serotec, UK), 5 g/ml mouse nti-bovine CD8 mAb (Serotec, UK) or 20 g/ml mouse nti-bovine CD21 mAb (Santa Cruz, USA) as a B-cell marker. n Alexa Fluor 488 (AF488) conjugated goat anti-mouse ross-absorbed secondary antibody (Invitrogen, Australia) as used to detect all reactive mAb antibodies (Table 1).

Techniques: