Journal: The Journal of Biological Chemistry

Article Title: Integrin β4 drives immune evasion and therapeutic resistance to PD-1 blockade in bladder cancer via MEK/ERK signaling

doi: 10.1016/j.jbc.2025.110941

Figure Lengend Snippet: ITGB4 positively correlated with enfortumab vedotin target NECTIN4. A , the RNA-Seq analysis in the GSE268855 dataset after overexpressing ITGB4 ; B , the correlation analysis in the DepMap dataset. p values are shown in the figure. C , Pan-cancer data analysis in the DepMap database revealed the correlation between ITGB4 and NECTIN4 . p values were shown in the figure. D – G , HT-1376 and 5637 cells were transfected with shITGB4 or shControl for 72 h. The harvested cells were used for Western blot analysis ( D ) and RT-qPCR detection ( E – G ). H – K , HT-1376 and 5637 cells were transfected with overexpressed ITGB4 plasmid or empty vector for 72 h, and harvested cells were used for Western blot analysis ( H ) and RT-qPCR detection ( I–K ). Data presents as mean ± SD with three replicates. ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. L , the tissue microarray of bladder cancer stained with ITGB4 and NECTIN4, respectively. The typical IHC images stained with ITGB4 and NECTIN4 are shown in the panel . M , the correlation of these two proteins is shown in the panel , and the p -value is indicated in the figure.

Article Snippet: The primary antibodies used were as follows: ITGB4 (#21738-1-AP, Proteintech, 1:500 dilution), p-ITGB4(Y1510) (#YP0755, Immunoway, 1:500 dilution), MEK-1(#YM8273, Immunoway, 1:1000 dilution), p-MEK-1 (T386) (#YP0425, Immunoway, 1:500 dilution), ERK1/2(#YM8336, Immunoway, 1:2000 dilution), p-ERK1/2 (T202/T204) (#YM8452, Immunoway, 1:2000 dilution), c-Jun (#YM8321, Immunoway, 1:2000 dilution), p-c-Jun (S63) (#28907-1-AP, Proteintech, 1:500 dilution), PD-L1(#28076-1-AP, Proteintech, 1:300 dilution), NECTIN4 (#21903-1-AP, Proteintech, 1:2000 dilution), HER2(#18299-1-AP, Proteintech, 1:2000 dilution), TROP2(#27360-1-AP, Proteintech, 1:500 dilution), GADPH (#10494-1-AP, Proteintech, 1:5000 dilution).

Techniques: RNA Sequencing, Transfection, Western Blot, Quantitative RT-PCR, Plasmid Preparation, Microarray, Staining

Journal: The Journal of Biological Chemistry

Article Title: Integrin β4 drives immune evasion and therapeutic resistance to PD-1 blockade in bladder cancer via MEK/ERK signaling

doi: 10.1016/j.jbc.2025.110941

Figure Lengend Snippet: ITGB4 plays a vital role in BLCA sequential therapies. A mechanism diagram depicting that the highly expressed ITGB4 in cisplatin-resistant BLCA activated the MEK/ERK pathway through tyrosine-1510 phosphorylation, upregulated the expression level of PD-L1 in BLCA, and caused anti-PD-1 treatment resistance. Meanwhile, the overexpressed ITGB4 was positively correlated to NECTIN4, promoting the sensitivity of BLCA to enfortumab vedotin treatment.

Article Snippet: The primary antibodies used were as follows: ITGB4 (#21738-1-AP, Proteintech, 1:500 dilution), p-ITGB4(Y1510) (#YP0755, Immunoway, 1:500 dilution), MEK-1(#YM8273, Immunoway, 1:1000 dilution), p-MEK-1 (T386) (#YP0425, Immunoway, 1:500 dilution), ERK1/2(#YM8336, Immunoway, 1:2000 dilution), p-ERK1/2 (T202/T204) (#YM8452, Immunoway, 1:2000 dilution), c-Jun (#YM8321, Immunoway, 1:2000 dilution), p-c-Jun (S63) (#28907-1-AP, Proteintech, 1:500 dilution), PD-L1(#28076-1-AP, Proteintech, 1:300 dilution), NECTIN4 (#21903-1-AP, Proteintech, 1:2000 dilution), HER2(#18299-1-AP, Proteintech, 1:2000 dilution), TROP2(#27360-1-AP, Proteintech, 1:500 dilution), GADPH (#10494-1-AP, Proteintech, 1:5000 dilution).

Techniques: Phospho-proteomics, Expressing

Journal: JCI Insight

Article Title: Nectin-4 reduces T cell effector function and is a therapeutic target in pancreatic cancer

doi: 10.1172/jci.insight.194290

Figure Lengend Snippet: ( A ) Representative images of CD155 (top) and Nectin-4 (bottom) IHC staining with low and high intensity. Scale bar: 100 μm. ( B ) Dot plot showing the distribution of immune reactive scores (IRS; CD155, n = 69; Nectin-4, n = 68). ( C ) Stacked columns depicting the proportion of patients with CD155 and Nectin-4 expression according to intensity. ( D ) Kaplan-Meier analysis of overall survival of R0-resected patients with PDAC according to low or high CD155 (left) or Nectin-4 (right) expression. P values of log rank test are indicated. ( E ) Table and forest plot depicting survival hazard ratios (HR) with 95% CI of CD155 and Nectin-4 IRS in multivariate Cox proportional hazards regression analysis including both R0- and R1-resected patients, shown as a function of clinicopathological parameters. * P < 0.05; ** P < 0.01.

Article Snippet: The membrane was incubated overnight at 4°C with Nectin-4 antibody (17402, Cell Signaling Technology, RRID:AB_2798785).

Techniques: Immunohistochemistry, Expressing

Journal: JCI Insight

Article Title: Nectin-4 reduces T cell effector function and is a therapeutic target in pancreatic cancer

doi: 10.1172/jci.insight.194290

Figure Lengend Snippet: ( A and B ) Percentage of T cells among all immune cells (left) and percentage of indicated T cell subsets among all T cells (right) in blood ( A ) and PDAC ( B ) for low and high Nectin-4 expression ( n = 68). ( C and D ) Percentage of TIGIT, CD226 and CD96 expression (from left to right) for indicated T cell subsets in blood ( C ) and PDAC ( D ) for low and high Nectin-4 expression. Each point represents data from 1 patient. Medians are shown as horizontal lines. Unpaired 2-tailed t tests with Holm-Šídák correction. * P < 0.05.

Article Snippet: The membrane was incubated overnight at 4°C with Nectin-4 antibody (17402, Cell Signaling Technology, RRID:AB_2798785).

Techniques: Expressing

Journal: JCI Insight

Article Title: Nectin-4 reduces T cell effector function and is a therapeutic target in pancreatic cancer

doi: 10.1172/jci.insight.194290

Figure Lengend Snippet: ( A ) IFN-γ and TNF-α production by peripheral T cells from patients with PDAC after in vitro stimulation with anti-CD3 and anti-CD28 in the presence of plate-bound (pb, n = 8) or soluble (s, n = 4) Nectin-4. Each point represents data from 1 patient. Data are shown as mean ± SD. Unpaired 2-tailed t tests with Welch’s correction. * P < 0.05; ** P < 0.001; *** P < 0.001; **** P < 0.0001. ( B ) Representative expression level of PVRL4 by qPCR (bars indicate mean of technical duplicates). ( C ) Nectin-4 expression in PDAC PDOs by Western blot. ( D ) Dose response curves from PDAC PDOs treated with FOLFIRINOX, gemcitabine plus paclitaxel (Gem/Pac), or enfortumab vedotin. The relative viability in percentage at a given drug concentration of 2 independent biological replicates is shown. ( E ) Z scores generated from relative AUC from dose response curves from PDAC PDOs either treated with FOLFIRINOX, Gem/Pac, or enfortumab vedotin. ( F ) Representative images of 2 PDAC PDOs either treated with the standard regimen FOLFIRINOX, Gem/Pac, or enfortumab vedotin. PDOs were stained with caspase-3 dye profiling apoptosis (green) and imaged after 3 days. Scale bar: 50 μm.

Article Snippet: The membrane was incubated overnight at 4°C with Nectin-4 antibody (17402, Cell Signaling Technology, RRID:AB_2798785).

Techniques: In Vitro, Expressing, Western Blot, Concentration Assay, Generated, Staining