Journal: Nature Communications

Article Title: Group I Paks are essential for epithelial- mesenchymal transition in an Apc -driven model of colorectal cancer

doi: 10.1038/s41467-018-05935-6

Figure Lengend Snippet: Alternative splicing of CD44 contributes to EMT in PID -expressed cells. a Real-time PCR analysis showed relative expression levels of standard isoform, total and variants of CD44 in PID + and PID - primary cells. Results were normalized to GAPDH . b Immunoblot analysis and quantification of CD44 isoform in PID + (4, 7, and 26) and PID - (35, 48, and 56) cells. GAPDH served as the internal control. c Immunoblot analysis showed protein levels and localization of hnRNPM, PCBP1, ESRP1, Snai1 and P-Snai1 in PID + (4 and 26) and PID - (35 and 56) primary cells. Lamin A/C and Vinculin served as loading control for cytoplasmic (C) and nuclear (N) fraction, respectively. Graphs represented quantification of proteins in nuclear and cytosolic fraction. d Immunoblot analysis and quantification of E-Cadherin, N-Cadherin, Zeb1, ESRP1, and Snai1 in PID - primary cells either transduced with retrovirus bearing empty vector (EV), wild-type Snai1 (WT) or virus expressing mutant Snai1-S246A (SA). GAPDH serve as a loading control. e Immunoblot analysis and quantification of ESRP1, CD44, E-Cadherin and GAPDH in PID + primary cells either transducted with sh-Scramble or shRNA against ESRP1. f Schematic illustration of invasiness inhibition by PID peptide

Article Snippet: Antibodies against β-Catenin (#610154; 1/2000), E-Cadherin (#610182; 1/1000) and N-Cadherin (#610920; 1/1000) were from BD Transduction Laboratories; Occludin (#sc-5562; 1/1000), PCBP1 (#sc-393076; 1/1000), CD44 (#sc-18849; 1/300), and Lamin A/C (#sc-7292; 1/1000) were from Santa Cruz Biotechnology; hnRNPM (#TA301557; 1/1000); ESRP1 (#NBP1-82201; 1/1000) was from Novus Biologicals; Snai1 (#ab180714; 1/500) and phospho-Snai1 (#ab63568; 1/250) were from Abcam.

Techniques: Alternative Splicing, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control, Transduction, Plasmid Preparation, Virus, Mutagenesis, shRNA, Inhibition

Journal: Nature Communications

Article Title: Septin5 deletion enhances β-cell exocytosis by releasing microtubule-tethered insulin granules onto plasma membrane

doi: 10.1038/s41467-025-57421-5

Figure Lengend Snippet: a Western blotting showing Septin5 is present in human and rodent islets. Rat insulinoma INS-1 is shown for comparison. b Western blotting showing Septin5 is present human islets, whose levels were not reduced in T2D human islets, as is also the case with GK rat islets in ( a ). SNARE proteins are reduced in both T2D human and GK islets (in ( a )) compared to normal islets. c Quantification of band intensity (normalized to tubulin) in ( b ) showing similar expression level of Septin5 in human normal and T2D islets ( n = 3 normal donors and 3 T2D donors). Statistical significance was assessed by a two-tailed Student’s t -test (n.s., not significant). For boxplots, boxes indicate 25th and 75th percentiles, the lines within boxes mark medians, whiskers above and below boxes indicate the 90th and 10th percentiles. d Lightning confocal images of human islet ( top ) and dispersed single β-cell ( bottom ) immunostained with Septin5 and insulin show Septin5 is abundant in pancreatic β-cells. A similar experiment was performed on mouse islets (Supplementary Fig. ). e Quantification of endogenous Septin5 by immunostaining showing the heterogeneity of Septin5 expression in both dispersed human β-cells and intact islets from six non-diabetic and three T2D donors. Data are normalized to the average intensity. All the data shown are representative of at least 3 separate experiments. Source data are provided as a file.

Article Snippet: Septin5 (catalog 11631-1-AP) and actin (catalog 66009-1-lg) antibodies were from Proteintech Group (Rosemont, IL, USA); tubulin antibody (catalog 11H10) was from Cell Signaling (Danvers, MA, USA); STX-1A (catalog SAB5600255) and insulin (catalog SAB4200691) antibodies were from Sigma-Aldrich (St. Louis, MO, USA); Munc18a antibody (catalog 610336) was from BD Transduction Laboratories (NULL, USA); SNAP25 antibody (catalog 111 011) was from Synaptic Systems GmbH (Goettingen, Germany).

Techniques: Western Blot, Comparison, Expressing, Two Tailed Test, Immunostaining

Journal: Nature Communications

Article Title: Septin5 deletion enhances β-cell exocytosis by releasing microtubule-tethered insulin granules onto plasma membrane

doi: 10.1038/s41467-025-57421-5

Figure Lengend Snippet: a – d IPGTTs were performed to assess blood glucose ( a , b ) and secreted insulin levels ( c , d ) and shown as AUC (area under the curve) analysis encompassing 120 min of the IPGTT for blood glucose ( b ) and insulin levels ( d ). N = 5 mice for each group, with summary graphs shown as mean ± SEM. Septin5 deletion was first confirmed in isolated islets, which did not affect the total islet insulin content (Supplementary Fig. ). For boxplots, boxes indicate 25th and 75th percentiles, the lines within boxes mark medians, whiskers above and below boxes indicate the 90th and 10th percentiles. e Blood glucose levels during IPITT showing no significant difference ( n = 4 for control, and 6 for Septin5 KO from three biologically independent experiments). f Total insulin content is not affected in Septin5 KO mice. g – h Biphasic GSIS determined by Islet perifusion assays ( g ) of WT vs Septin5 KO mice and the corresponding AUC analysis ( h ) for first phase (first 10 min) and second phase (next 20 min) GSIS. Summary graphs from 4 biologically independent experiments shown as mean ± SEM. i Insulin-immunostained pancreatic sections. Septin5 KO does not alter the size, number and distribution of islets within the pancreas. Supplementary Fig. 2b, c shows the analysis for islet size and number. Scale bars, left , 1 mm, right , 100 µm. Statistical significance was assessed by a two-tailed Student’s t -test (* p < 0.05, the precise statistical results indicated in figures). Source data are provided as a file.

Article Snippet: Septin5 (catalog 11631-1-AP) and actin (catalog 66009-1-lg) antibodies were from Proteintech Group (Rosemont, IL, USA); tubulin antibody (catalog 11H10) was from Cell Signaling (Danvers, MA, USA); STX-1A (catalog SAB5600255) and insulin (catalog SAB4200691) antibodies were from Sigma-Aldrich (St. Louis, MO, USA); Munc18a antibody (catalog 610336) was from BD Transduction Laboratories (NULL, USA); SNAP25 antibody (catalog 111 011) was from Synaptic Systems GmbH (Goettingen, Germany).

Techniques: Isolation, Control, Two Tailed Test

Journal: Nature Communications

Article Title: Septin5 deletion enhances β-cell exocytosis by releasing microtubule-tethered insulin granules onto plasma membrane

doi: 10.1038/s41467-025-57421-5

Figure Lengend Snippet: a Western blotting showed that Septin5 level was 81.9% reduced compared to control (Ad-control) when human islets infected with Ad-Septin5 shRNA (Septin5 KD) ( n = 3. Quantitative analysis of the experiments shown in Supplementary Fig. ). b Human islets perifusion assay showed that depletion of Septin5 in human islets increased both first-and second-phase GSIS. c AUC analysis of first-phase (10–20 min) and second-phase (21–41 min) GSIS. Basal secretion was not significantly affected. Plotted are the mean values of 4 biologically independent experiments, and bars represent SEM for ( b , c ). d – g TIRF imaging showed the fusion events in the first phase (first 5 min) and second phase (6–15 min) ( n = 10 cells for each condition from 5 normal donors. Supplementary Movie ), expressed as mean ± SEM. f The summary of fusion events with pre-docked and newcomer granules in first and second phase in both conditions. Pre-docked granules at basal (2.8 mM glucose) were not different between WT and Septin5 KO β-cells (Supplementary Fig. ). Boxes indicate 25th and 75th percentiles, the lines within boxes mark medians, whiskers above and below boxes indicate the 90th and 10th percentiles. g Normalized cumulative fusion events of SGs per unit area from control and Septin5 KD human β-cells. Statistical significance was assessed by a two-tailed Student’s t -test (* p < 0.05, the precise statistical results indicated in figures). Source data are provided as a Source Data file.

Article Snippet: Septin5 (catalog 11631-1-AP) and actin (catalog 66009-1-lg) antibodies were from Proteintech Group (Rosemont, IL, USA); tubulin antibody (catalog 11H10) was from Cell Signaling (Danvers, MA, USA); STX-1A (catalog SAB5600255) and insulin (catalog SAB4200691) antibodies were from Sigma-Aldrich (St. Louis, MO, USA); Munc18a antibody (catalog 610336) was from BD Transduction Laboratories (NULL, USA); SNAP25 antibody (catalog 111 011) was from Synaptic Systems GmbH (Goettingen, Germany).

Techniques: Western Blot, Control, Infection, shRNA, Imaging, Two Tailed Test

Journal: Nature Communications

Article Title: Septin5 deletion enhances β-cell exocytosis by releasing microtubule-tethered insulin granules onto plasma membrane

doi: 10.1038/s41467-025-57421-5

Figure Lengend Snippet: a Septin5 depletion in human islets increases GSIS by affecting MT rather than actin microfilaments. Insulin secretion over 30 min upon 16.7 mM glucose stimulation as measured by ELISA. Human islets after two-round infection with control or Ad-Septin5 shRNA were incubated with DMSO, nocodazole, latrunculin or taxol. Data are presented as box-plots where the central line represents the median value, the box shows percentiles 25–75 and the whiskers include the most extreme values (two-tailed Student’s t -test, n.s. denotes not significant). b – e Representative 3D structured illumination microscopy images of human β-cells infected with granule marker NPY-EGFP and Ad-shRNA-Control ( b , c ) or Ad-shRNA-Septin5 ( d , e ) for 48 h and then immunostained with tubulin. Pseudo-color encodes the depth. Color corresponds to the depth from the coverslip surface, with red being closest to the PM surface and Cyan being in cell interior. b 3D reconstructed Super-resolution imaging shows Septin5 is assembled at MT-PM contact sites ( bottom , white arrows, higher magnification view of white dashed rectangular region) as rod and punctate structures, some granules were observed anchoring to Septin5-rich sites along the MT ( bottom , white circles, higher magnification view of white dashed rectangular region). c The TIRF optical section images of 3D SIM highlighted the MT-PM contact sites of the region as the dashed box indicated in ( b ). d Septin5 deletion does not alter the MT meshwork or the anchoring of granules ( right , white circles). Image is depth coded as indicated. e The TIRF optical section images of 3D SIM highlighted the MT-PM contact sites of the region as the dashed box indicated in ( d ). Representative results of three independent experiments.

Article Snippet: Septin5 (catalog 11631-1-AP) and actin (catalog 66009-1-lg) antibodies were from Proteintech Group (Rosemont, IL, USA); tubulin antibody (catalog 11H10) was from Cell Signaling (Danvers, MA, USA); STX-1A (catalog SAB5600255) and insulin (catalog SAB4200691) antibodies were from Sigma-Aldrich (St. Louis, MO, USA); Munc18a antibody (catalog 610336) was from BD Transduction Laboratories (NULL, USA); SNAP25 antibody (catalog 111 011) was from Synaptic Systems GmbH (Goettingen, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Control, shRNA, Incubation, Two Tailed Test, Microscopy, Marker, Imaging

Journal: Nature Communications

Article Title: Septin5 deletion enhances β-cell exocytosis by releasing microtubule-tethered insulin granules onto plasma membrane

doi: 10.1038/s41467-025-57421-5

Figure Lengend Snippet: a , b Spatiotemporal analysis of SG dynamics during glucose stimulation in WT ( a ) and Septin5-depleted ( b ) human β-cells by TIRFM. Representative TIRFM images ( left ), sum of SG trajectories ( right ), and time-lapse images of typical SG dynamics before fusion ( bottom ). c , d Histogram of the granule diffusion coefficient D measured from the trajectory of individual granules in WT ( c ) and Septin5 knockdown ( d ) human β-cell after glucose stimulation. Diffusion coefficient D is a measure of how constrained diffusion of the particles, smaller values indicate more restricted movement. The raw data (dots) are fitted with a three-component Gaussian function (red line), corresponding to the nearly immobile, restricted and highly dynamic SG population, respectively (from left to right ). n = 6 cells, ~700 trajectories analyzed. e The spatial profile of SGs “exploring” the PM before fusion showed that Septin5 depletion increases SGs access to PM. To visualize the “exploring” region before fusion, time-lapse images of ~70 fusion events spatially aligned by the center coordinates of fusion site within a window of size 9×9 pixels were temporally averaged during the 5-s preceding fusion pore opening (aligning point illustrated on left ). Spatial profiles of the average signal of NPY-EGFP at fusion sites showed in middle , single-granule profile included as reference. Dashed lines represent raw data, and the histogram was fitted with Gaussian function ( right ). Width, FWHM. f Septin5 depletion decreased the stability of MT after high glucose stimulation as indicated by the ratio ( fire ) of glu-tubulin ( turquoise ) to total tubulin ( vermillion ). The higher magnification ( right ) of regions as the dashed box ( left ) indicated showed example of region with decreased MT stability ( bottom ) compared to control ( top ). Representative images of human islets treated with high glucose for 30 min and then stained for stable MT marker, Glu-tubulin and total MT marker tubulin ( n = 3 independent experiments, quantitative analysis of the experiments shown in Supplementary Fig. ).

Article Snippet: Septin5 (catalog 11631-1-AP) and actin (catalog 66009-1-lg) antibodies were from Proteintech Group (Rosemont, IL, USA); tubulin antibody (catalog 11H10) was from Cell Signaling (Danvers, MA, USA); STX-1A (catalog SAB5600255) and insulin (catalog SAB4200691) antibodies were from Sigma-Aldrich (St. Louis, MO, USA); Munc18a antibody (catalog 610336) was from BD Transduction Laboratories (NULL, USA); SNAP25 antibody (catalog 111 011) was from Synaptic Systems GmbH (Goettingen, Germany).

Techniques: Diffusion-based Assay, Knockdown, Control, Staining, Marker

Journal: Nature Communications

Article Title: Septin5 deletion enhances β-cell exocytosis by releasing microtubule-tethered insulin granules onto plasma membrane

doi: 10.1038/s41467-025-57421-5

Figure Lengend Snippet: a Normal and T2D human β-cell infected with granule marker NPY-tdOrange2 and loaded with Ca 2+ sensor 2 μM Fluo5F as well as EGTA-AM stimulated with high K + (75 mM). The localized Fluo5F signal indicated the Ca 2+ influx sites after depolarization. b , c Fusion events of granules associated with L-type Ca 2+ channels in human normal ( b ) and T2D ( c ) β-cells. d , e Septin5 depletion improves GSIS in human T2D islets. The corresponding area-under-the-curve (AUC) analysis was summarized in ( e ) (Data from n = 3 biologically independent experiments were represented as mean ± SEM). f – g The glucose-stimulated insulin secretion of GK rat islets is assessed by perfusion. 25 islets exposed to 2.8 mM glucose for 10 min, followed by 16.7 mM glucose for 30 min, during which the sample collected every 2.5 min. The corresponding AUC analysis was summarized in ( g ) (Data from n = 3 biologically independent experiments were represented as mean ± SEM). h – k Septin5 deletion increases Ca 2+ sensitivity to both low ( h , i ) and high ( j , k ) calcium concentrations but did not cause any change in calcium channel opening or density (Supplementary Fig. ). h The whole-cell capacitance change recorded during direct application of 200 nM free Ca 2+ via patch pipette into human β-cells. i The average capacitance increments in control and Septin5 KD cells at 150 s following initiation of Ca 2+ infusion and normalized to initial cell size. j Exocytosis was elicited by flash photolysis and monitored by whole-cell membrane capacitance measurement. Averaged capacitance changes from control (black, n = 11) and Septin5 KD (gray, n = 11) cells. Arrow indicates the flash time. k Averaged amplitudes of the highly calcium-sensitive pool from control (black) and Septin5 KD (white) cells respectively ( n = 11 from three independent experiments). Statistical significance was assessed by a two-tailed Student’s t -test (the precise statistical results indicated in figures). For boxplots, boxes indicate 25th and 75th percentiles, the lines within boxes mark medians, whiskers above and below boxes indicate the 90th and 10th percentiles. Source data are provided as a file.

Article Snippet: Septin5 (catalog 11631-1-AP) and actin (catalog 66009-1-lg) antibodies were from Proteintech Group (Rosemont, IL, USA); tubulin antibody (catalog 11H10) was from Cell Signaling (Danvers, MA, USA); STX-1A (catalog SAB5600255) and insulin (catalog SAB4200691) antibodies were from Sigma-Aldrich (St. Louis, MO, USA); Munc18a antibody (catalog 610336) was from BD Transduction Laboratories (NULL, USA); SNAP25 antibody (catalog 111 011) was from Synaptic Systems GmbH (Goettingen, Germany).

Techniques: Infection, Marker, Transferring, Control, Membrane, Two Tailed Test

Journal: Heliyon

Article Title: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

doi: 10.1016/j.heliyon.2018.e00545

Figure Lengend Snippet: PD-1H expression after transfection of PD-1H overexpression vector or siPD-1H vector. (A) Gating strategy. (B) Isotype stained monocytes. (C) Empty vector Neon-transfected monocytes. (D) PD-1H overexpressing vector transfected monocytes. (E) siScramble transfected monocytes. (F) siPD-1H transfected monocytes. (G) Related ratio of PD-1H expression after vector vs. PD-1H transfection using average MFIs from three different experiments represented in C and D. (H) Related ratio of PD-1H expression after siScramble vs. siPD-1H transfection using average MFIs from three different experiments represented in E and F.

Article Snippet: After generation of sepsis mice for 24 h, the blood was collected to analyze the PD-1H expression in monocytes by staining the ficoll-isolated human PBMCs with anti-human PD-1H antibody (R&D systems, Inc.), as well as human CD45, CD3, CD4, CD8, and CD14 antibodies.

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Staining

Journal: Heliyon

Article Title: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

doi: 10.1016/j.heliyon.2018.e00545

Figure Lengend Snippet: Gene sets enriched in PD-1H overexpressed monocytes, analyzed by KEGG (Kyoto Encyclopedia of Genes and Genomes) gene sets.

Article Snippet: After generation of sepsis mice for 24 h, the blood was collected to analyze the PD-1H expression in monocytes by staining the ficoll-isolated human PBMCs with anti-human PD-1H antibody (R&D systems, Inc.), as well as human CD45, CD3, CD4, CD8, and CD14 antibodies.

Techniques: Ubiquitin Proteomics, Infection, Phospho-proteomics, Transduction

Journal: Heliyon

Article Title: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

doi: 10.1016/j.heliyon.2018.e00545

Figure Lengend Snippet: Gene sets enriched in PD-1H overexpressed monocytes, analyzed by Biocarta gene sets.

Article Snippet: After generation of sepsis mice for 24 h, the blood was collected to analyze the PD-1H expression in monocytes by staining the ficoll-isolated human PBMCs with anti-human PD-1H antibody (R&D systems, Inc.), as well as human CD45, CD3, CD4, CD8, and CD14 antibodies.

Techniques:

Journal: Heliyon

Article Title: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

doi: 10.1016/j.heliyon.2018.e00545

Figure Lengend Snippet: Gene set enrichment analysis of Biocarta pathways. Genes were ranked according to the difference between average transcript expression of PD-1H overexpressed cases and controls. Each pathway in Biocarta database was then assigned the (normalized) enrichment score using GSEA (version 2.2.4). Three enrichment plots with highest enrichment scores are shown for representatively.

Article Snippet: After generation of sepsis mice for 24 h, the blood was collected to analyze the PD-1H expression in monocytes by staining the ficoll-isolated human PBMCs with anti-human PD-1H antibody (R&D systems, Inc.), as well as human CD45, CD3, CD4, CD8, and CD14 antibodies.

Techniques: Expressing

Journal: Heliyon

Article Title: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

doi: 10.1016/j.heliyon.2018.e00545

Figure Lengend Snippet: Global gene expression difference between PD-1H overexpressed cases and vectors. Two-way unsupervised clustering was performed on gene expression profiles of PD-1H overexpressed cases and vectors. Green and red color indicate relatively lower/higher expression of genes, respectively, while black indicates intermediated gene expression. Distance in the dendrogram tree was calculated using Ward's method.

Article Snippet: After generation of sepsis mice for 24 h, the blood was collected to analyze the PD-1H expression in monocytes by staining the ficoll-isolated human PBMCs with anti-human PD-1H antibody (R&D systems, Inc.), as well as human CD45, CD3, CD4, CD8, and CD14 antibodies.

Techniques: Gene Expression, Expressing

Journal: Heliyon

Article Title: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

doi: 10.1016/j.heliyon.2018.e00545

Figure Lengend Snippet: Summary of the most significant differentially regulated genes/gene clusters in PD-1H overexpressed monocytes.

Article Snippet: After generation of sepsis mice for 24 h, the blood was collected to analyze the PD-1H expression in monocytes by staining the ficoll-isolated human PBMCs with anti-human PD-1H antibody (R&D systems, Inc.), as well as human CD45, CD3, CD4, CD8, and CD14 antibodies.

Techniques: Activation Assay

Journal: Heliyon

Article Title: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

doi: 10.1016/j.heliyon.2018.e00545

Figure Lengend Snippet: IPA analysis of upstream transcription regulators in PD-1H overexpressed monocytes. IPA was used to identify the transcription regulators with significant target gene overlap with changes in PD-1H overexpressed monocytes transcriptome. The histogram showed the most significant activators and inhibitors using a Z-score cut-off of ≥2 (for activators) or ≤-2 (for inhibitors).

Article Snippet: After generation of sepsis mice for 24 h, the blood was collected to analyze the PD-1H expression in monocytes by staining the ficoll-isolated human PBMCs with anti-human PD-1H antibody (R&D systems, Inc.), as well as human CD45, CD3, CD4, CD8, and CD14 antibodies.

Techniques:

Journal: Heliyon

Article Title: Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile

doi: 10.1016/j.heliyon.2018.e00545

Figure Lengend Snippet: PD-1H expression in sham-treated or sepsis mice. (A) Representative PD-1H expression level from one of the sham-treated or sepsis mouse. (B) Average percentages (left histogram) and MFI (right histogram) of PD-1H expression. CLP: C ecal L igation and P uncture.

Article Snippet: After generation of sepsis mice for 24 h, the blood was collected to analyze the PD-1H expression in monocytes by staining the ficoll-isolated human PBMCs with anti-human PD-1H antibody (R&D systems, Inc.), as well as human CD45, CD3, CD4, CD8, and CD14 antibodies.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Clinico-histopathological characteristics of the 136 cutaneous melanomas according to CD33, VISTA and PD-1 expression.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Correlations among CD33, VISTA, and PD-1 expression.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Expression of CD33 and VISTA in cutaneous melanoma. ( A ) High intratumoral expression (immunohistochemical score of 3, red color, cytoplasmic) of CD33 in acral melanoma (× 100). ( B ) High intratumoral expression (score 3, red color, cytoplasmic) of VISTA in nodular melanoma (× 100). ( C ) High peritumoral expression (score 3, red color, cytoplasmic) of CD33 in acral melanoma (× 100). ( D ) High peritumoral expression (score 3, red color, cytoplasmic) of VISTA in nodular melanoma (× 200). ( E ) Dual immunofluorescence staining of ( A ) CD33 and VISTA (CD33, red, cytoplasmic; VISTA, green, cytoplasmic) and ( F ) PD-1 and VISTA (PD-1, red, cytoplasmic; VISTA, green, cytoplasmic). Double-positive cells are shown in yellow (white arrow). VISTA, V-domain Ig suppressor of T-cell activation; PD-1, programmed cell death protein-1.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing, Immunohistochemical staining, Immunofluorescence, Staining, Activation Assay

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Patient survival according to CD33 and VISTA expression. ( A ) OS according to CD33 expression. ( B ) OS according to VISTA expression. ( C ) Worse OS in patients with high expression of both CD33 and VISTA. ( D ) OS according to the degree of CD33 and VISTA expression ( E ) Worse OS in cases with dual expression of VISTA and PD-1 among patients with high expression of PD-1. VISTA, V-domain Ig suppressor of T-cell activation; PD-1, programmed cell death protein-1.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing, Activation Assay

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Univariate and multivariate analyses of OS and PFS.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing