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: Cell Death & Disease

Article Title: Targeting RNA polymerase I to boost natural killer cell anticancer activity in multiple myeloma

doi: 10.1038/s41419-025-08196-6

Figure Lengend Snippet: A Cell surface expression of HLA-E was assessed by flow cytometry on SKO-007(J3) cells treated with CX-5461in the absence or in the presence of the ATRi (AZD6738/Ceralasertib, 1 µM) for 48 h. Histograms depict the mean fluorescence intensity (MFI) of HLA-E, after subtracting the MFI of the isotype control (* P < 0.05). B Western Blot analysis of HLA-E in SKO-007(J3) cells untreated or treated with the indicated inhibitors for 48 h. Expression of p85 was used as protein loading control. A representative Western Blot is shown with densitometric analysis normalized to p85, together with the quantification of three independent experiments (* P < 0.05). C NK cell degranulation against CX-treated SKO, in the presence or absence of ATRi was evaluated as the percentage CD107a. PBMCs from healthy donors served as effector cells. Data represent the mean value from four independent experiments (* P < 0.05). D Western Blot analysis of pS6K in SKO-007(J3) cells untreated or treated with RNA Pol I inhibitors for 24 h. A representative Western Blot is shown coupled to densitometric analysis, normalized to β-Actin. E Total mRNA was obtained from SKO-007(J3) cells stably infected with lentiviruses pLKO-shRNA-CBP20 or pLKO non-targeting shRNA (control) and analyzed for CBP20 mRNA expression by Real-Time qRT-PCR. Data were normalized with GAPDH and referred to the cells infected with non-target shRNA, considered as calibrator. F , G HLA-E and HLA-ABC expression were analyzed by flow cytometry on SKO-007(J3) pLKO non-target shRNA or pLKO-shRNA-CBP20 infected cells, treated with CX-5461 as described above. The white colored histograms represent basal expression of the indicated HLA, while grey histograms represent the expression after drug treatment. Data are the average of four independent experiments (* P < 0.05). H , I HLA-E expression was analyzed by flow cytometry on SKO-007(J3) and RPMI-8226 cells treated with CX-5461 in the presence of the LMP7/PSMB8 inhibitor M3258 (300 nM). Data are the average of three independent experiments (* P < 0.05).

Article Snippet: The highly selective immunoproteasome subunit LMP7 inhibitor M3258, was acquired from MedChemExpress (New Jersey, USA).

Techniques: Expressing, Flow Cytometry, Fluorescence, Control, Western Blot, Stable Transfection, Infection, shRNA, Quantitative RT-PCR