Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 1 Immunoproteasome expression profile in muscle-invasive bladder cancer (MIBC). A Expression of immunoproteasome subunits PSMB8, PSMB9 and PSMB10 in pan-tumor tissues and corresponding normal tissues derived from the TCGA dataset. Red boxes outline upregulation of PSMB8, PSMB9 and PSMB10 in bladder cancer tissues in contrast to normal tissues. Data are expressed as individual spots per subject with mean of the logarithm of transcripts per million. *p < 0.05, **p < 0.01, ***p < 0.001. B Expression of PSMB8, PSMB9 and PSMB10 in MIBC (n = 369) and normal tissues (n = 19) from the TCGA cohort. Data are expressed as individual spots per subject with mean ± SEM of the logarithm of transcripts per million. P-values are indicated in each graph. C Immunohistochemistry staining scores of PSMB8, PSMB9 and PSMB10 in MIBC (n = 67) and normal tissues (n = 18) derived from the CQUCH cohort. Data are expressed as individual spots per subject with mean ± SEM of each group. P-values are indicated in each graph. D Representative positive and negative immunohistochemistry stainings of PSMB8, PSMB9 and PSMB10 in MIBC and normal tissues derived from the CQUCH cohort. Scale bar: 40 μm. Positive stainings are divided into low and high expression by an average immunohistochemistry staining score

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing, Derivative Assay, Immunohistochemistry, Staining

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 2 Association of the expression of immunoproteasome subunits with survival prognosis in MIBC patients. A Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients in the CQUCH cohort with high and low expression of PSMB8, PSMB9 and PSMB10. B Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients in the TCGA cohort with high and low expression of PSMB8, PSMB9 and PSMB10

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 3 Association of the expression of immunoproteasome subunits with survival prognosis in MIBC patients receiving immunotherapy. A Kaplan–Meier curve of overall survival for MIBC patients receiving immune checkpoint inhibitor treatment in the IMvigor210 cohort with high and low expression of PSMB8, PSMB9 and PSMB10. B Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients receiving immune checkpoint inhibitor treatment in the CQUCH cohort with high and low expression of PSMB8, PSMB9 and PSMB10

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 4 Effect of immunotherapy (IMT) on survival prognosis in MIBC patients with different expression levels of immunoproteasome subunits. A Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients receiving IMT or not receiving (non-IMT) immune checkpoint inhibitor treatment in the CQUCH cohort with high expression of PSMB8, PSMB9 and PSMB10. B Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients receiving IMT or not receiving (non-IMT) immune checkpoint inhibitor treatment in the CQUCH cohort with low expression of PSMB8, PSMB9 and PSMB10

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 6 Association of immunoproteasome subunits with inflammatory factors in MIBC. Expression correlation heatmap of the association of IFN-γ and TNF with the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 in MIBC of the TCGA cohort (A) and the IMvigor210 cohort (B). Correlation coefficient between each two molecules is written in each square. Color bar on the right side of each heatmap shows the correlation coefficient. Positive correlation is shown in red, negative and blue. C, D Kaplan–Meier curve of overall survival of MIBC patients receiving immune checkpoint inhibitor treatment in the IMvigor210 cohort with high and low expression of IFN-γ or TNF

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 7 Association of immunoproteasome subunits with tumor infiltrating immune cells in MIBC. Correlation lollipop charts of tumor infiltrating immune cells which were associated with the expression of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 in MIBC of the TCGA cohort (A) and the IMvigor210 cohort (B). Lollipop size shows the correlation coefficient between each infiltrating immune cell type and each immunoproteasome subunit. P values were labeled on the right side of each chart. Values of P < 0.05 were considered statistically significant and marked in red

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing, Labeling

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 8 Association of immunoproteasome subunits with different immune-related functions and signaling in MIBC. A GO analysis of the association of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 with immune-related functions in MIBC of the TCGA cohort. Color bar shows the p-value on the right side of each chart. The number of related genes were shown as the length of the column of each chart. B KEGG analysis of the association of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 with immune-related signaling in MIBC of the TCGA cohort. Color bar shows the p-value on the right side of each chart. The number of related genes were shown by dot size. C GSEA analysis of the association of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 with different immune cell functions in MIBC of the TCGA cohort

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques:

Journal: Biochemistry

Article Title: Differential Interactions of the Proteasome Inhibitor PI31 with Constitutive and Immuno-20S Proteasomes.

doi: 10.1021/acs.biochem.3c00707

Figure Lengend Snippet: Figure 3. PI31 inhibition of 20Si is attenuated compared to that of 20Sc. PI31 inhibition of latent and z-YA-activated 20Sc and 20Si activity was compared in multiple assays of proteasome function. Panel A. Effect of PI31 on latent and z-YA-activated hydrolysis of Suc-LLVY-AMC (upper) and casein (lower). Mean values for rates of substrate hydrolysis for triplicate assays were determined as described under Materials and Methods. Values for activity in the absence of z-YA and PI31 were set to 100% (control), and other values were calculated as a percentage of control, ± SD. Differences among results of different assay conditions were analyzed by 3-way ANOVA. Pair-wise differences were analyzed posthoc by Tukey’s HSD. PI31 significantly inhibited z-YA-activated 20Sc- and 20Si-catalyzed hydrolysis of both substrates (20Sc, p < 0001; 20Si, p = 0.0193). PI31 inhibition of 20Si was significantly attenuated compared to that of 20Sc for both substrates (p < 0.0001, denoted by *). Panel B. z-YA-activated 20Sc and 20Si hydrolysis of α-synuclein was determined in the presence (●) or absence (O) of PI31 (20S, 150 nM; PI31, 500 nM). At indicated times, samples were subjected to SDS-PAGE and stained with Coomassie Blue R-250. α-Synuclein content was quantified using Image Studio software (LiCOR). Values at time 0 were set to 100%, and other values within each group are represented as a percentage. Panel C. Me4BodipyFL-Ahx3Leu3-VS labeling of 20Sc and 20Si in the presence (●) or absence (O) of 5 mM z-YA and/or PI31. Labeling was conducted as described in Materials and Methods and quantified using Image Studio software (LiCOR). In each of the 3 independent experiments (shown as individual data points), labeling of each subunit in the absence of PI31 was set to 100%. Other values within that group are represented as percentages of that value. The means ± SD of the three independent experiments are shown. Panel D. Purified 20Sc and 20Si were assayed for hydrolysis of Suc-LLVY-AMC and casein in the presence or absence of 5 mM z-YA at indicated concentrations of PI31. Mean values for rates of hydrolysis of triplicate assays in the absence of PI31 were set to 100%, and other values were determined as a percentage of that value. Differences in activity between 20Sc and 20Si were analyzed by repeated measures of 2-way ANOVA. Pair-wise, concentration-matched differences in activity between 20Sc and 20Si were analyzed by Šídák’s multiple comparisons test, and significant differences are indicated at p ≤0.05 by (*) for the respective PI31 concentrations. Similar results were obtained in four independent experiments.

Article Snippet: Western blotting was performed as previously described.40 Proteins transferred to nitrocellulose membranes were blocked and blotted using primary antibodies against 6xHis (mouse monoclonal, Invitrogen MA1−135); 20S α4 (PSMA7) (mouse monoclonal antihuman, Enzo PW8120); PI31 C-terminus residues 241−271 (rabbit polyclonal antihuman, Enzo BML-PW9710); and 20S β2i (PSMB10) (rabbit polyclonal antihuman, Cell Signaling Technology, 78385S).

Techniques: Inhibition, Activity Assay, Control, SDS Page, Staining, Software, Labeling, Purification, Concentration Assay

Journal: Biochemistry

Article Title: Differential Interactions of the Proteasome Inhibitor PI31 with Constitutive and Immuno-20S Proteasomes.

doi: 10.1021/acs.biochem.3c00707

Figure Lengend Snippet: Figure 8. Structural determinants of PI31 for complex formation with 20Sc and 20Si. 20Sc or 20Si were preincubated with the indicated PI31 proteins and subjected to glycerol density gradient centrifugation as described under Materials and Methods. Panel A. Distribution of His-PI31 was determined by Western blotting using anti-His tag antibody. Control gradients were performed for the proteasome in the absence of PI31 using an antibody against the α4 subunit common to both 20Sc and 20Si and against PI31 in the absence of 20S. Panel B. Quantification of bands from Panel A. Similar results were obtained in three independent experiments.

Article Snippet: Western blotting was performed as previously described.40 Proteins transferred to nitrocellulose membranes were blocked and blotted using primary antibodies against 6xHis (mouse monoclonal, Invitrogen MA1−135); 20S α4 (PSMA7) (mouse monoclonal antihuman, Enzo PW8120); PI31 C-terminus residues 241−271 (rabbit polyclonal antihuman, Enzo BML-PW9710); and 20S β2i (PSMB10) (rabbit polyclonal antihuman, Cell Signaling Technology, 78385S).

Techniques: Gradient Centrifugation, Western Blot, Control

Journal: Biochemistry

Article Title: Differential Interactions of the Proteasome Inhibitor PI31 with Constitutive and Immuno-20S Proteasomes.

doi: 10.1021/acs.biochem.3c00707

Figure Lengend Snippet: Figure 10. Model of differential PI31 binding to, inhibition of, and hydrolysis by 20Sc and 20Si proteasomes. The intrinsically disordered C-terminal domain of PI31 enters the central chamber of 20Sc and 20Si proteasomes. PI311−271 binds to βc catalytic sites to inhibit their function while escaping hydrolysis itself. Truncated PI31s with C-termini that cannot reach the catalytic sites PI311−<193 feature no 20S inhibition and have reduced binding. PI311−271 also enters the central chamber of 20Si but interacts with βi catalytic subunits in a manner that leads to reduced proteasome inhibition and PI31 hydrolysis. For clarity, the images depict a single molecule of PI31 interacting with three of the six catalytic subunits of 20S proteasomes. However, two molecules of PI31 can enter the proteasome from opposite ends of the cylinder and interact with all six catalytic subunits.

Article Snippet: Western blotting was performed as previously described.40 Proteins transferred to nitrocellulose membranes were blocked and blotted using primary antibodies against 6xHis (mouse monoclonal, Invitrogen MA1−135); 20S α4 (PSMA7) (mouse monoclonal antihuman, Enzo PW8120); PI31 C-terminus residues 241−271 (rabbit polyclonal antihuman, Enzo BML-PW9710); and 20S β2i (PSMB10) (rabbit polyclonal antihuman, Cell Signaling Technology, 78385S).

Techniques: Binding Assay, Inhibition

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 1 Immunoproteasome expression profile in muscle-invasive bladder cancer (MIBC). A Expression of immunoproteasome subunits PSMB8, PSMB9 and PSMB10 in pan-tumor tissues and corresponding normal tissues derived from the TCGA dataset. Red boxes outline upregulation of PSMB8, PSMB9 and PSMB10 in bladder cancer tissues in contrast to normal tissues. Data are expressed as individual spots per subject with mean of the logarithm of transcripts per million. *p < 0.05, **p < 0.01, ***p < 0.001. B Expression of PSMB8, PSMB9 and PSMB10 in MIBC (n = 369) and normal tissues (n = 19) from the TCGA cohort. Data are expressed as individual spots per subject with mean ± SEM of the logarithm of transcripts per million. P-values are indicated in each graph. C Immunohistochemistry staining scores of PSMB8, PSMB9 and PSMB10 in MIBC (n = 67) and normal tissues (n = 18) derived from the CQUCH cohort. Data are expressed as individual spots per subject with mean ± SEM of each group. P-values are indicated in each graph. D Representative positive and negative immunohistochemistry stainings of PSMB8, PSMB9 and PSMB10 in MIBC and normal tissues derived from the CQUCH cohort. Scale bar: 40 μm. Positive stainings are divided into low and high expression by an average immunohistochemistry staining score

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing, Derivative Assay, Immunohistochemistry, Staining

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 2 Association of the expression of immunoproteasome subunits with survival prognosis in MIBC patients. A Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients in the CQUCH cohort with high and low expression of PSMB8, PSMB9 and PSMB10. B Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients in the TCGA cohort with high and low expression of PSMB8, PSMB9 and PSMB10

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 3 Association of the expression of immunoproteasome subunits with survival prognosis in MIBC patients receiving immunotherapy. A Kaplan–Meier curve of overall survival for MIBC patients receiving immune checkpoint inhibitor treatment in the IMvigor210 cohort with high and low expression of PSMB8, PSMB9 and PSMB10. B Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients receiving immune checkpoint inhibitor treatment in the CQUCH cohort with high and low expression of PSMB8, PSMB9 and PSMB10

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 4 Effect of immunotherapy (IMT) on survival prognosis in MIBC patients with different expression levels of immunoproteasome subunits. A Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients receiving IMT or not receiving (non-IMT) immune checkpoint inhibitor treatment in the CQUCH cohort with high expression of PSMB8, PSMB9 and PSMB10. B Kaplan–Meier curve of progression-free survival (upper panel) and overall survival (lower panel) for MIBC patients receiving IMT or not receiving (non-IMT) immune checkpoint inhibitor treatment in the CQUCH cohort with low expression of PSMB8, PSMB9 and PSMB10

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 6 Association of immunoproteasome subunits with inflammatory factors in MIBC. Expression correlation heatmap of the association of IFN-γ and TNF with the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 in MIBC of the TCGA cohort (A) and the IMvigor210 cohort (B). Correlation coefficient between each two molecules is written in each square. Color bar on the right side of each heatmap shows the correlation coefficient. Positive correlation is shown in red, negative and blue. C, D Kaplan–Meier curve of overall survival of MIBC patients receiving immune checkpoint inhibitor treatment in the IMvigor210 cohort with high and low expression of IFN-γ or TNF

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 7 Association of immunoproteasome subunits with tumor infiltrating immune cells in MIBC. Correlation lollipop charts of tumor infiltrating immune cells which were associated with the expression of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 in MIBC of the TCGA cohort (A) and the IMvigor210 cohort (B). Lollipop size shows the correlation coefficient between each infiltrating immune cell type and each immunoproteasome subunit. P values were labeled on the right side of each chart. Values of P < 0.05 were considered statistically significant and marked in red

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: Expressing, Labeling

Journal: Journal of translational medicine

Article Title: Immunoproteasome subunits are novel signatures for predicting efficacy of immunotherapy in muscle invasive bladder cancer.

doi: 10.1186/s12967-025-06207-w

Figure Lengend Snippet: Fig. 8 Association of immunoproteasome subunits with different immune-related functions and signaling in MIBC. A GO analysis of the association of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 with immune-related functions in MIBC of the TCGA cohort. Color bar shows the p-value on the right side of each chart. The number of related genes were shown as the length of the column of each chart. B KEGG analysis of the association of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 with immune-related signaling in MIBC of the TCGA cohort. Color bar shows the p-value on the right side of each chart. The number of related genes were shown by dot size. C GSEA analysis of the association of the immunoproteasome subunits PSMB8, PSMB9 and PSMB10 with different immune cell functions in MIBC of the TCGA cohort

Article Snippet: Following incubation in phosphate-buffered saline containing 10% species-appropriate normal serum to block non-specific binding at room temperature for 1 h, sections were incubated in a humidified chamber with primary antibodies against PSMB8 (1:100; proteintech, Wuhan, China), PSMB9 (1:100; proteintech), PSMB10 (1:100; proteintech) and PD-L1 (1:100; proteintech) using isotype-matched IgGs as negative controls at 4 °C overnight.

Techniques: