Journal: Oncoimmunology

Article Title: SERPINB9 is commonly amplified and high expression in cancer cells correlates with poor immune checkpoint blockade response

doi: 10.1080/2162402X.2022.2139074

Figure Lengend Snippet: SERPINB9 is the major differentially expressed gene in T cell-resistant vs. sensitive NSCLC cell lines. a) Schematic representation of the NSCLC tumor cell line panel in co-culture with T cells. Tumor cells were transduced with vectors encoding HLA-A02 and MART-1, while T cells were transduced with a MART-1-specific TCR. b) Tumor cells and T cells were co-cultured at a ratio of 1:1 for 16 hours, fixed and stained with crystal violet. Each data point indicates an individual biological replicate. Mean with S.D. c) Cell viability measurement by Cell titer blue (CTB) of the four most resistant versus the four most sensitive NSCLC cell lines treated with the indicated cytokines (100μg/ml) for 5 days. Unpaired t test was used for statistical analysis. *p<0.05 **p<0.001 d) Volcano plot showing differential expression analysis comparing the four most resistant versus the four most sensitive cell lines. Dashed lines indicate significance thresholds (FDR < 0.01, log2 fold change < -2 or > 2). e) Correlation of SERPINB9 mRNA normalized expression with viability measured as described in A and B. Each data point indicates an individual cell line. A simple linear regression test was used for statistical analysis. p= 0.0002. f) SERPINB9 protein quantification by western blot in the four most resistant versus four most sensitive cell lines. HSP90 serves as a loading control.

Article Snippet: The following antibodies were used: anti-SERPINB9 (PA5-101879, Thermo Fisher), HSP90 (4874S, CST) and anti-rabbit IgG (Thermo Fisher, G-21234).

Techniques: Co-Culture Assay, Transduction, Cell Culture, Staining, Quantitative Proteomics, Expressing, Western Blot, Control

Journal: Oncoimmunology

Article Title: SERPINB9 is commonly amplified and high expression in cancer cells correlates with poor immune checkpoint blockade response

doi: 10.1080/2162402X.2022.2139074

Figure Lengend Snippet: Genetic ablation of SERPINB9 sensitizes tumor cells to T cell killing. a) Western blot showing SERPINB9 protein expression in sgRNA control (Ctrl)- or sgSERPINB9-expressing LCLC-103H and A549 T cell-resistant cell lines. HSP90 serves as a loading control. b) Cell apoptosis measurement by a Caspase3/7 dye in a co-culture of sgCtrl- or sgSERPINB9-expressing tumor cells (LCLC-103H left, A549 right) with or without T cells, as indicated. Number of Caspase3/7-positive cells were imaged by an Incucyte system every 2 hours. Mean with S.D. 2-way ANOVA test was used for statistical analysis. ****p<0.0001. The small right-hand panels show the end point measurements of four independent biological experiments with different T cell donors. Unpaired t test was used for statistical analysis. ****p<0.0001. c) Cell viability measurement by crystal violet staining of endpoints in B) at different T cell: tumor cell ratios.

Article Snippet: The following antibodies were used: anti-SERPINB9 (PA5-101879, Thermo Fisher), HSP90 (4874S, CST) and anti-rabbit IgG (Thermo Fisher, G-21234).

Techniques: Western Blot, Expressing, Control, Co-Culture Assay, Staining

Journal: Oncoimmunology

Article Title: SERPINB9 is commonly amplified and high expression in cancer cells correlates with poor immune checkpoint blockade response

doi: 10.1080/2162402X.2022.2139074

Figure Lengend Snippet: SERPINB9 overexpression causes resistance to T cell killing. a) Western blot showing SERPINB9 protein expression in cells overexpressing (ORF for Open Reading Frame) GFP or SERPINB9 in T cell-sensitive cell lines EBC-1 (left) and NCI-H358 (right). HSP90 serves as a loading control. b) Cell apoptosis measurement by a Caspase3/7 dye in a co-culture of GFP- or SERPINB9-expressing tumor cells (EBC-1 left, NCI-H358 right) with or without T cells, as indicated. Number of Caspase3/7 positive cells were measured by Incucyte analysis every 2 hours. Mean with S.D. 2-way ANOVA test was used for statistical analysis. ****p<0.0001. The small right-hand panels show the end point measurements of four independent biological experiments with different T cell donors. Unpaired t test was used for statistical analysis. ****p<0.0001. c) Cell viability measurement by Crystal violet staining of endpoints in E) at different T cell: tumor cell ratios.

Article Snippet: The following antibodies were used: anti-SERPINB9 (PA5-101879, Thermo Fisher), HSP90 (4874S, CST) and anti-rabbit IgG (Thermo Fisher, G-21234).

Techniques: Over Expression, Western Blot, Expressing, Control, Co-Culture Assay, Staining

Journal: Endocrine

Article Title: Effects and plasma proteomic analysis of GLP-1RA versus CPA/EE, in combination with metformin, on overweight PCOS women: a randomized controlled trial

doi: 10.1007/s12020-023-03487-4

Figure Lengend Snippet: ELISA validation of plasma candidate protein marker levels. Plasma levels of PRDX6 ( A ), GSTO1 ( B ), GSTP1 ( C ), GSTM1 ( D ), FN1 ( E ) and SERPINB9 ( F ) were measured in the CPA/EE+Met and GLP-1RA+Met groups at baseline and post treatment. n = 27–30. * P value < 0.05, ** P value < 0.01, *** P value < 0.001, paired t test

Article Snippet: The validation analysis of the differential plasma proteins was performed by ELISA, following the manuals of corresponding ELISA kits for GSTP1 (ELH-GSTP1-1, Raybiotech, Georgia, USA), GSTM2 (P81730, Cusabio, Wuhan, China), GSTO1 (Abbexa, abx151761, TX 77042, USA), PRDX6 (SEF756Hu, Cloudclone, Wuhan, China), Fibronectin (FN1) (ab108848, Abcam, Cambridge, USA), and SERPINB9 (SED390Hu, Cloudclone, Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Clinical Proteomics, Marker