Journal: Current Therapeutic Research, Clinical and Experimental

Article Title: The Berberine Derivative BBR684 Inhibits VDAC Oligomerization to Suppress Ferroptosis in Acute Kidney Injury

doi: 10.1016/j.curtheres.2026.100825

Figure Lengend Snippet: BBR684 attenuates Erastin-induced ferroptosis in HK-2 cells by suppressing lipid peroxidation and activating the antioxidant response. (A) Flow cytometric analysis of lipid peroxidation using the C11-BODIPY probe in HK-2 cells treated as indicated for 12 hours. Data are presented as mean fluorescence intensity (MFI). (B) Intracellular Fe²⁺ levels measured by a ferrous iron colorimetric assay. (C, D) Levels of malondialdehyde (MDA, C) and reduced glutathione (GSH, D) in cell lysates. (E, F) Representative immunofluorescence images (E) and quantification (F) of 4-hydroxynonenal (4-HNE) adducts (red) in HK-2 cells. Nuclei were counterstained with DAPI (blue). Scale bar, 50 µm. (G) Western blot analysis of key ferroptosis-related proteins: glutathione peroxidase 4 (GPX4), heme oxygenase-1 (HO-1), and nuclear factor erythroid 2–related factor 2 (NRF2). β-actin served as the loading control. Data are from 3 independent biological replicates (n = 3) and presented as mean ± SEM. Significance was determined by one-way ANOVA followed by Tukey's HSD post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001; n.s., not significant.

Article Snippet: Antibodies against β-Actin (Catalog #GB15003-100) were from Servicebio (Wuhan, China).

Techniques: Fluorescence, Colorimetric Assay, Immunofluorescence, Western Blot, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: BPGM as an intrinsic brake to constrain metastasis through phospho-epigenetic-mediated carnitine biosynthesis suppression

doi: 10.1016/j.neo.2026.101299

Figure Lengend Snippet: Overexpression of BPGM inhibits tumor metastasis in vitro and in vivo . (A) Silencing BPGM promoted migration of tumor cells. Tumor cells stably expressing shBPGM and its control cells (shCtrl) were examined. Scale bar, 200 μm. (B) Overexpressing BPGM suppressed migration of tumor cells. Tumor cells stably expressing BPGM and its control cells (Ctrl) were examined. Scale bar, 200 μm. (C) The mRNA levels of MMP2 and MMP9 reduced in BPGM-overexpressing tumor cells. SK-HEP-1 cells stably expressing BPGM (BPGM-OE) and its ctrl cells were employed to detect the mRNA levels of MMP2 and MMP9 by qPCR analysis. β-actin was used as an internal control. (D-E) Xenografts of stable Bpgm-overexpressing cells displayed a lower rate of liver and lung metastasis and less metastatic nodules in the liver. For (D), Hepa-Ctrl ( n = 7) and Hepa-BPGM sublines ( n = 6) were inoculated under the capsule of the left hepatic lobe of C57BL/6 mice. Upper panel, a schematic diagram of orthotopic hepatic implantation model. The number of metastatic rate and nodules is shown (D, lower panel). Scale bar, 1 cm. Hematoxylin-eosin staining was performed on serial sections of livers (E, left panel) and lungs (E, right panel) to detect the metastatic nodules. The red arrows indicated the metastatic nodules. Scale bar in left panel, 200 µm; Scale bar in right panel, 100 μm. (F-H) Overexpressing of BPGM inhibited lung metastasis of tumor xenografts. Scale bar in F, 1 cm. B16-F10 cells transfected with Ctrl ( n = 6) or BPGM-OE ( n = 6) was injected into the tail vein of C57BL/6 mice, respectively. Upper panel in F, a schematic diagram of lung metastasis model by tail vein injection. H&E staining of lung sections was performed to observe metastatic foci (G). For G, scale bar in left panel, 500 μm; scale bar in right panel, 200 μm. The number of melanoma nodules and lung metastasis is shown in H. (I) The model deciphers the inhibitory role of BPGM in tumor metastasis. Error bar: mean ± SEM. P -values are labeled above the bar chart.

Article Snippet: The antibodies used included mouse antibody against β-actin (BM0627, Boster, Wuhan, China), rabbit antibody against BPGM (17173-1-AP, Proteintech), EZH2 (F0281, Selleck), phospho-EZH2 (Thr345) (TA3584S, Abmart, Shanghai, China), phospho-CDK1 (Thr14) (AP1465, Abclonal, Wuhan, China), ubiquitin (10201-2-AP, Proteintech), HIF1α (36169, Cell Signaling Technology, CST, Beverly, MA, USA), H3K4me3 (91264, Active Motif), H3K79me3 (cat 49-1020, Thermos Fisher), H3K9me3 (61014, Active Motif), H3K27me3 (91168, Active Motif) and Histone 3 (F0057, Selleck).

Techniques: Over Expression, In Vitro, In Vivo, Migration, Stable Transfection, Expressing, Control, Staining, Transfection, Injection, Labeling

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: Survival analysis of patients based on key genes and their correlation with APM genes (A–D) Survival curves of groups with high and low expression of key genes ( GPX3 , CAMK2N2 , TRIM36 , and RND2 ) (George’s cohort ). (E) Scatterplot of the average expression of key genes vs. the average correlation with antigen presentation genes. (F and G) Scatterplots of the correlation between TRIM36, CAMK2N2 and HLA-A, HLA-B, HLA-C, B2M, respectively. Statistical significance was defined as p < 0.05. Statistical analyses were performed using R software. Survival analysis was conducted using the Kaplan-Meier method, and differences between groups were compared using the log rank test. Correlations were analyzed using Pearson’s correlation analysis. HLA, human leukocyte antigen; B2M, beta-2 microglobulin.

Article Snippet: After dewaxing the sections, they were blocked with goat serum, then incubated with primary antibodies against B2M (Boster: A00456-2, Servicebio: GB21303) and β-catenin (Proteintech: Cat#66379-1-Ig).

Techniques: Expressing, Immunopeptidomics, Software

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: TRIM36 and CAMK2N2 promote tumor growth in vivo , inhibit the expression of MHC I, and suppress CD8 + T cell activation in SCLC (A) qRT-PCR detection of the relative expression levels of Camk2n2 gene in the NC group and Sh Camk2n2 group, and Trim36 gene in the NC group and Sh Trim36 group of RP1 cells. qRT-PCR data were normalized to β-actin (ACTB) expression. (B) Gross morphology of subcutaneous tumors in the NC group, Sh Camk2n2 group, and Sh Trim36 group after dissection on the 24th day when mice were injected with cells ( n = 5 per group). (C) Statistics of tumor weights in each group. (D) Curves of tumor volume changes over time in each group. (E and F) Flow cytometry detection and statistics of the expression of H2D/H2K and B2M in tumors. (G–I) Flow cytometry analysis of the proportion of CD3 + CD8 + T cells and IFNγ + CD8 + T cells. Cell experiments were independently repeated 3 times ( n = 3). Data are presented as mean ± SD. Statistical significance: ∗∗ p < 0.01, ∗∗∗ p < 0.001. Statistical analyses were performed using GraphPad Prism software. Multiple group comparisons were conducted using one-way analysis of variance (one-way ANOVA) followed by Bonferroni correction for pairwise comparisons. NC, negative control; MFI, mean fluorescence intensity.

Article Snippet: After dewaxing the sections, they were blocked with goat serum, then incubated with primary antibodies against B2M (Boster: A00456-2, Servicebio: GB21303) and β-catenin (Proteintech: Cat#66379-1-Ig).

Techniques: In Vivo, Expressing, Activation Assay, Quantitative RT-PCR, Dissection, Injection, Flow Cytometry, Software, Negative Control, Fluorescence

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: TRIM36 and CAMK2N2 inhibit the expression of antigen presentation genes in SCLC cells (A and B) The relative expression levels of H2D , H2K , B2m , and Tap1 genes in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group of RP1 cells with or without IFNγ treatment detected by qRT-PCR. qRT-PCR data were normalized to ACTB expression. (C and D) The relative expression levels of Psmb8 , Psmb9 , and Nlrc5 genes in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group with or without IFNγ treatment detected by qRT-PCR. qRT-PCR data were normalized to ACTB expression. (E and F) WB detection of the expression levels of HLA-ABC, B2M, and ACTB proteins in the untreated group, NC group, Sh CAMK2N2 group, and Sh TRIM36 group of H1688 cells with or without IFNγ treatment. WB band intensities were quantified using ImageJ software (normalized to ACTB). (G–J) MFI of H2D/H2K and B2M in the untreated group, NC group, Sh Camk2n2 group, and Sh Trim36 group with or without IFNγ treatment in RP1 cells detected by flow cytometry. All cell experiments were independently repeated three times ( n = 3). Data are presented as mean ± SD. Statistical significance definitions: n.s., no significance; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Statistical analyses were performed using GraphPad Prism software. Multiple group comparisons were conducted using one-way ANOVA followed by Bonferroni correction. NC, negative control; HLA, human leukocyte antigen; B2M, beta-2 microglobulin; H2D/H2K, histocompatibility 2-D region antigen/H2-K region antigen; MFI, mean fluorescence intensity.

Article Snippet: After dewaxing the sections, they were blocked with goat serum, then incubated with primary antibodies against B2M (Boster: A00456-2, Servicebio: GB21303) and β-catenin (Proteintech: Cat#66379-1-Ig).

Techniques: Expressing, Immunopeptidomics, Quantitative RT-PCR, Software, Flow Cytometry, Negative Control, Fluorescence

Journal: Frontiers in Plant Science

Article Title: Peroxynitrite-mediated tyrosine nitration modulates β-1,3-glucanase activity and potato defense against Phytophthora infestans

doi: 10.3389/fpls.2026.1796014

Figure Lengend Snippet: Analysis of β-1,3-glucanase gene expression, protein accumulation, and enzymatic activity in leaves of potato genotypes challenged with Phytophthora infestans . Transcript levels determined by qRT-PCR in resistant (A) and susceptible (B) potato inoculated with P. infestans . Protein accumulation in leaves of the resistant (C) and susceptible (D) genotypes following challenge inoculation with the pathogen. Equal amounts of protein (25 µg per lane) were loaded onto SDS–PAGE gels, and Coomassie Brilliant Blue (CBB) staining was used as a loading control. Enzymatic activity of β-1,3-glucanases was measured spectrophotometrically using laminarin as substrate in resistant (E) and susceptible (F) potato after inoculation. Values represent the mean ± SD of at least three independent experiments (n = 9). Asterisks indicate values that differ significantly from mock-inoculated (control) potato leaves at *P < 0.05.

Article Snippet: After transfer, the membranes were blocked (3% BSA) and incubated with polyclonal antibodies against β-1,3-glucanase (Agrisera) at a 1:1000 dilution.

Techniques: Gene Expression, Activity Assay, Quantitative RT-PCR, SDS Page, Staining, Control

Journal: Frontiers in Plant Science

Article Title: Peroxynitrite-mediated tyrosine nitration modulates β-1,3-glucanase activity and potato defense against Phytophthora infestans

doi: 10.3389/fpls.2026.1796014

Figure Lengend Snippet: Effect of SIN-1 (ONOO – donor) on recombinant β-1,3-glucanase activity (A) , protein accumulation and tyrosine nitration (B) . Samples were incubated at varying concentrations of SIN-1 (0.05–2 mM). Equal amounts of recombinant protein (1 µg per lane) were loaded onto the SDS-PAGE gel. Coomassie Brilliant Blue (CBB) staining served as a loading control.

Article Snippet: After transfer, the membranes were blocked (3% BSA) and incubated with polyclonal antibodies against β-1,3-glucanase (Agrisera) at a 1:1000 dilution.

Techniques: Recombinant, Activity Assay, Nitration, Incubation, SDS Page, Staining, Control

Journal: Frontiers in Plant Science

Article Title: Peroxynitrite-mediated tyrosine nitration modulates β-1,3-glucanase activity and potato defense against Phytophthora infestans

doi: 10.3389/fpls.2026.1796014

Figure Lengend Snippet: Structure of potato β-1,3-glucanase. The tertiary structure was generated by homology modeling, based on the coordinates of β-1,3-glucanase from Hevea brasiliensis (PDB ID: 4HPG, chain A). Surface active pocket identification by CASTp server: (A) Light Green, Red, and light yellow color boxes highlight the amino acid residues present in the binding site. (B) Shows the Tyr 59 and Tyr 203 in the catalytic pocket of β-1,3-glucanase. Calculated distances between the two Tyr residues (in Å) and the catalytic acid (E120) or the nucleophile (E265) (C, D) . Yellow dashed lines indicate distances between the residues.

Article Snippet: After transfer, the membranes were blocked (3% BSA) and incubated with polyclonal antibodies against β-1,3-glucanase (Agrisera) at a 1:1000 dilution.

Techniques: Generated, Binding Assay

Journal: Frontiers in Plant Science

Article Title: Peroxynitrite-mediated tyrosine nitration modulates β-1,3-glucanase activity and potato defense against Phytophthora infestans

doi: 10.3389/fpls.2026.1796014

Figure Lengend Snippet: Evolutionary conservation of amino acid residues of potato β-1,3-glucanase (accession number P52401 ) was analyzed using the ConSurf server. Each residue was assigned a conservation grade ranging from 1 (variable) to 9 (highly conserved) based on a multiple-sequence alignment of homologous proteins. Residues located in the catalytic pocket are marked in black frames (A) . Characterization of the six highly conserved tyrosine (Y) residues (conservation score 9/9) present in potato β-1,3-glucanase in terms of amino acid content within a nine-residue window of linear sequence around the tyrosine (Primary structure), solvent-accessible surface area (ASA), secondary-structure organization, and residues within 5 Å from the two carbons susceptible to nitration. Residues meeting the nearby-residue criterion (basic amino acids in the immediate vicinity of the tyrosine residue within the primary sequence and/or the acidic residues within 5 Å from the atom susceptible to nitration) are shown in bold (B) .

Article Snippet: After transfer, the membranes were blocked (3% BSA) and incubated with polyclonal antibodies against β-1,3-glucanase (Agrisera) at a 1:1000 dilution.

Techniques: Residue, Sequencing, Solvent, Nitration