Journal: Clinical and Experimental Medicine

Article Title: Multi-omics identification of a programmed cell death-related signature and potential target P4HB for bladder cancer based on a 101-combination machine learning and experimental validation

doi: 10.1007/s10238-026-02146-y

Figure Lengend Snippet: Multiple machine learning algorithms were integrated to screen BLCA signature genes. (A) 10 different machine learning algorithms were incorporated to identify the genetic markers based on these 21 PCDGs. (B-D) 12 signature genes were identified by the CoxBoost algorithm and GBM algorithm. (E) The heat map revealed a significant upregulation of P4HB, ATP13A2, TFRC, TRAF7, and RRP12 in the high-risk group sample. (F) Patients with BLCA in the high-risk group had significantly lower OS compared to those in the low-risk group. (G) ROC curve showed excellent predictive accuracy for 1-, 3-, and 5-year interval risk scores in the TCGA-BLCA cohort

Article Snippet: After blocked, the membranes were then incubated overnight at 4 °C with primary antibodies targeting P4HB (dilution 1:1000, Cat. #3501, CST, USA).

Techniques:

Journal: Clinical and Experimental Medicine

Article Title: Multi-omics identification of a programmed cell death-related signature and potential target P4HB for bladder cancer based on a 101-combination machine learning and experimental validation

doi: 10.1007/s10238-026-02146-y

Figure Lengend Snippet: Pseudotime trajectory analysis of P4HB in tumor cells using CytoTRACE2 and Monocle2. ( A ) Tumor cells were extracted and clustered into 14 distinct subgroups. (B) Expression of P4HB across the 14 tumor subgroups. (C) The differentiation status of tumor cells was visualized using CytoTRACE2, with colors representing the degree of differentiation. (D) The box plot shows the predicted ordering of tumor cell subpopulations by CytoTRACE2. (E-F) Pseudotime trajectory of tumor cells colored by pseudotime (E) and distribution of tumor cells (F). (G) Expression of P4HB across pseudotime. (H) Slingshot trajectory inference showing three distinct lineages originating from stem-like clusters. (I) Dynamic expression patterns of P4HB along each inferred lineage

Article Snippet: After blocked, the membranes were then incubated overnight at 4 °C with primary antibodies targeting P4HB (dilution 1:1000, Cat. #3501, CST, USA).

Techniques: Expressing

Journal: Clinical and Experimental Medicine

Article Title: Multi-omics identification of a programmed cell death-related signature and potential target P4HB for bladder cancer based on a 101-combination machine learning and experimental validation

doi: 10.1007/s10238-026-02146-y

Figure Lengend Snippet: Multiple analyses indicating a significant association between P4HB and the tumorigenesis as well as ROS pathway in BLCA. (A) Violin plot shows the differential expression of 12 hub genes between high- and low-risk score groups. (B) Boxplots show the overall expression levels of the 12 hub genes between cancer and normal tissues in the TCGA-BLCA cohort. (C) Paired difference analysis of 12 hub genes in the TCGA-BLCA cohort. (D) Radar map of P4HB expression levels in pan-cancer. (E) Immunohistochemical staining results from HPA database showed the expression of P4HB. (F) Risk regression analysis of P4HB in multiple bladder cancer cohorts (G) Differential expression analysis of P4HB in multiple progressive bladder cancer cohorts. (H) Differential expression analysis of P4HB in bladder cancer samples of different stages. (I) Expression density of P4HB at the single-cell level and cellular distribution of ROS pathway score

Article Snippet: After blocked, the membranes were then incubated overnight at 4 °C with primary antibodies targeting P4HB (dilution 1:1000, Cat. #3501, CST, USA).

Techniques: Quantitative Proteomics, Expressing, Immunohistochemical staining, Staining, Single Cell

Journal: Clinical and Experimental Medicine

Article Title: Multi-omics identification of a programmed cell death-related signature and potential target P4HB for bladder cancer based on a 101-combination machine learning and experimental validation

doi: 10.1007/s10238-026-02146-y

Figure Lengend Snippet: P4HB expression and function in BLCA. (A) RT-qPCR results of P4HB mRNA expression in BLCA tissues from 25 paired adjacent normal and tumor samples. (B-C) P4HB mRNA (B) and protein (C) expression in BLCA cell lines (mean ± SD, n = 3, *** p < 0.001). (D-E) The interference efficiency of P4HB shRNA in BIU87 (D) and UMUC3 (E) was evaluated by RT-qPCR and western blot (mean ± SD, n = 3). (F-H) CCK-8 (F), colony assays (G), and flow cytometry (H) determined that blocking P4HB inhibited BLCA cells’ proliferation and growth, but promoted cells apoptosis (mean ± SD, n = 3, *** p < 0.001)

Article Snippet: After blocked, the membranes were then incubated overnight at 4 °C with primary antibodies targeting P4HB (dilution 1:1000, Cat. #3501, CST, USA).

Techniques: Expressing, Quantitative RT-PCR, shRNA, Western Blot, CCK-8 Assay, Flow Cytometry, Blocking Assay

Journal: International Dental Journal

Article Title: A Novel DSPP Mutation in Dentinogenesis Imperfecta Shields Type II: Clinical, Genetic and Stem Cell Perspectives

doi: 10.1016/j.identj.2025.100937

Figure Lengend Snippet: Measurement of multi-lineage capacity of SHEDs from DGI patients and controls. (A-D) mRNA expression levels of DSPP, RUNX2, OCN and ALP in 4 groups of SHEDs. DGI represents the experimental group, while control1, control2 and control3 represent different control groups. Data are expressed as mean ± SD, with * P < .05 indicating statistical significance. (E) Protein levels of DSPP, RUNX2, OCN and ALP of DGI and Control SHEDs were presented as the GAPDH normalised ratio by western-blotting. Data are expressed as mean ± SD, with * P < .05 indicating statistical significance. (F) Differences in calcium deposition ability among SHEDs from one experimental group (DGI) and 3 control groups (Control1, Control2 and Control3) (40×). After alizarin red staining, red calcium nodules were visible under an inverted microscope. Bar graph showing the calcium deposition area after alizarin red staining in the 4 groups following osteogenic induction. Data are expressed as mean ± SD, with * P < .05 indicating statistical significance. (G) Adipogenic differentiation results of SHEDs from one experimental group (DGI) and 3 control groups (Control1, Control2 and Control3). (H) Effect of DSPP c.2590delG variant on protein localisation. Confocal microscopy shows intracellular localisation of the GFP tagged WT and frameshift mutant (c.2590delG) DSPP in odontoblast. Anti-CANX antibody was used for ER staining. Nuclei were stained with DAPI. The DSPP c.2590delG was found predominantly in the ER.

Article Snippet: The slides were incubated overnight at 4°C with rabbit polyclonal anti-calnexin (CANX) antibody (cat: A0692, ABClonal) at a 1:600 dilution, followed by a 2-hour incubation with Alexa Fluor 594-conjugated goat anti-rabbit IgG(H+L) (cat: AS039, ABClonal).

Techniques: Expressing, Control, Western Blot, Staining, Inverted Microscopy, Variant Assay, Confocal Microscopy, Mutagenesis