Journal: Bioactive Materials

Article Title: A multimodal ROS logic-gated therapeutic platform disrupts the vicious cycle of senescence to promote aged bone defect repair

doi: 10.1016/j.bioactmat.2026.02.002

Figure Lengend Snippet: Transcriptomic and molecular analysis of the potential pathways involved in MMBOx-mediated BMSCs rejuvenation. (A) Circular heatmap showing differentially expressed genes (DEGs) associated with cell senescence, inflammation, and osteogenesis in senescent BMSCs treated with MMBOx@GPP compared to GPP. (B) Gene Ontology (GO) enrichment analysis of upregulated DEGs. (C) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of upregulated DEGs. (D) Gene Set Enrichment Analysis (GSEA) plots of the glutathione metabolic process (ES: enrichment score; NES: normalized enrichment score; FDR: false discovery rate). (E) Heatmap of DEGs enriched in aging-related GO terms. (F) Western blot analysis of Keap1, Nrf2, Nqo1, Gclc, and GAPDH protein expression in BMSCs. (G) Quantitative analysis of protein band intensities ( n = 3). (H) Representative flow cytometry plots of ThiolTracker™ fluorescence staining indicating intracellular glutathione levels. (I) Quantification of intracellular GSH/GSSG ratio in BMSCs ( n = 3). (J) Schematic diagram illustrating the proposed mechanism by which MMBOx attenuates BMSCs senescence via Nrf2 pathway activation and glutathione metabolism enhancement. Data are expressed as mean ± SD; ∗ P < 0.05, ∗∗∗ P < 0.001.

Article Snippet: Western blotting was employed to evaluate protein expression of key targets, including Keap1 (Affinity, AF5266; 1:1000), Nrf2 (Proteintech, 16396-1-AP; 1:1000), Nqo1 (Abcam, ab80588; 1:10,000), Gclc (Proteintech, 12601-1-AP; 1:25000) and GAPDH (Proteintech, 60004-1-Ig; 1:50,000).

Techniques: Western Blot, Expressing, Flow Cytometry, Fluorescence, Staining, Activation Assay

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 GPX4 (Catalog #3F5G5), HO-1 (Catalog #10701-1-AP), NRF2 (Catalog #16396-1-AP), VDAC (Catalog #10866-1-AP), Cleaved Caspase-3 (Catalog #68773-1-Ig), and CD45 (Catalog #98035-1-RR) were purchased from Proteintech.

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

Journal: Microbiology Spectrum

Article Title: Verapamil HCl demonstrates antiviral activity against porcine reproductive and respiratory syndrome virus by regulating Ca 2+ influx and promoting type I interferon production

doi: 10.1128/spectrum.03280-25

Figure Lengend Snippet: Verapamil HCl upregulates Nrf2/HO-1 expression to suppress PRRSV replication. ( A–C ) PAMs were treated with various concentrations of Verapamil HCl (10, 20, 40, and 80 µM) or cobalt protoporphyrin (CoPP) (40 µM) for 24 h and then harvested to extract RNA and protein. Nrf2, HO-1, and Keap1 expression was further analyzed using qPCR and western blot. ( D–F ) PAMs were treated with various concentrations of Verapamil HCl (10, 20, 40, and 80 µM) or CoPP (40 µM) for 2 h. Subsequently, these cells were subjected to PRRSV SD16 infection (MOI = 0.1) for 1 h and then harvested to detect HO-1, Nrf2, and Keap1 expression at the mRNA and protein levels using qPCR and western blot. ( G and H ) PAMs were transfected with siHO-1/siNrf2 or siNC and then infected with PRRSV SD16 in the presence or absence of Verapamil HCl (80 µM) for 24 h. HO-1, Nrf2, and ORF-7 expression levels were tested using qPCR and western blot. GAPDH served as the internal control, and β-actin served as the loading control. The data shown are representative of three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001: compared with 0 µM Verapamil HCl-treated cells.

Article Snippet: The protein bands were transferred to polyvinylidene difluoride (PVDF) membranes, which were blocked with 5% skimmed milk powder for 1 h. Subsequently, the membranes were incubated with the indicated primary antibodies, including anti-PRRSV-1 and PRRSV-2 N protein antibodies; anti-heme oxygenase 1 (HO-1) mouse mAb (Servicebio); anti-p38/p-p38 rabbit pAb (CST, MA, USA); anti-ERK1/2 and p-ERK1/2 rabbit pAbs (Servicebio); anti-JNK1 + JNK2 + JNK3 rabbit pAb (Servicebio); anti-p-JNK mouse antibody (Santa Cruz, CA, USA), anti-Nrf2 antibody (Servicebio), anti-Keap1 rabbit pAb (Servicebio), anti-Lamin B1 rabbit pAb (Servicebio), and anti-β-actin antibody (Abcam, Cambridge, UK).

Techniques: Expressing, Western Blot, Infection, Transfection, Control

Journal: Microbiology Spectrum

Article Title: Verapamil HCl demonstrates antiviral activity against porcine reproductive and respiratory syndrome virus by regulating Ca 2+ influx and promoting type I interferon production

doi: 10.1128/spectrum.03280-25

Figure Lengend Snippet: Verapamil HCl activates the p38/Nrf2/HO-1 cell signaling pathway. ( A ) MARC-145 cells were incubated with Verapamil HCl (80 µM) and collected at different time points (0, 30, 60, and 120 min) to detect the phosphorylated and total protein levels of p38, ERK1/2, and JNK using western blot. MARC-145 cells were incubated with a mixture of Verapamil HCl and a JNK inhibitor (SP600125), ERK1/2 inhibitor (PD98059), or p38 inhibitor (SB203580) for 2 h, and these cells were harvested to analyze Nrf2 ( B ) and HO-1 ( C ) expression using qPCR. ( D ) MARC-145 cells were pretreated with 80 µM Verapamil HCl for 2 h and then infected with 0.1 MOI PRRSV SD16 for 1 h. The protein expression levels of Nrf2 in the nuclear and cytoplasmic fractions were analyzed using western blot at 24 hpi. Lamin B1 and β-actin served as the loading controls. ( E ) Meanwhile, the amount of p-38, p-p38, and N protein expression in the total cellular fraction was also determined using western blot. MARC-145 cells were incubated with a mixture of Verapamil HCl (80 µM) and a p38 inhibitor (SB203580) for 2 h prior to infection with PRRSV SD16 (MOI = 0.1). The Nrf2 and N protein expression in these cells was analyzed at 24 hpi using qPCR ( F ) and western blot ( G ). GAPDH served as the internal control, and β-actin acted as the loading control. All the data shown are representative of three independent experiments. ** P < 0.01 and *** P < 0.001.

Article Snippet: The protein bands were transferred to polyvinylidene difluoride (PVDF) membranes, which were blocked with 5% skimmed milk powder for 1 h. Subsequently, the membranes were incubated with the indicated primary antibodies, including anti-PRRSV-1 and PRRSV-2 N protein antibodies; anti-heme oxygenase 1 (HO-1) mouse mAb (Servicebio); anti-p38/p-p38 rabbit pAb (CST, MA, USA); anti-ERK1/2 and p-ERK1/2 rabbit pAbs (Servicebio); anti-JNK1 + JNK2 + JNK3 rabbit pAb (Servicebio); anti-p-JNK mouse antibody (Santa Cruz, CA, USA), anti-Nrf2 antibody (Servicebio), anti-Keap1 rabbit pAb (Servicebio), anti-Lamin B1 rabbit pAb (Servicebio), and anti-β-actin antibody (Abcam, Cambridge, UK).

Techniques: Incubation, Western Blot, Expressing, Infection, Control

Journal: Microbiology Spectrum

Article Title: Verapamil HCl demonstrates antiviral activity against porcine reproductive and respiratory syndrome virus by regulating Ca 2+ influx and promoting type I interferon production

doi: 10.1128/spectrum.03280-25

Figure Lengend Snippet: Schematic diagram depicting the anti-PRRSV effects of Verapamil HCl. Verapamil HCl activates AMPK by promoting the expression of p-p38, which activates Nrf2 and its downstream gene HO-1. HO-1 upregulation promotes the host cellular type I IFN response and inhibits the production of pro-inflammatory factors. Meanwhile, Verapamil HCl reverses the calcium ion imbalance induced by PRRSV. Collectively, these mechanisms inhibit PRRSV infection at multiple stages.

Article Snippet: The protein bands were transferred to polyvinylidene difluoride (PVDF) membranes, which were blocked with 5% skimmed milk powder for 1 h. Subsequently, the membranes were incubated with the indicated primary antibodies, including anti-PRRSV-1 and PRRSV-2 N protein antibodies; anti-heme oxygenase 1 (HO-1) mouse mAb (Servicebio); anti-p38/p-p38 rabbit pAb (CST, MA, USA); anti-ERK1/2 and p-ERK1/2 rabbit pAbs (Servicebio); anti-JNK1 + JNK2 + JNK3 rabbit pAb (Servicebio); anti-p-JNK mouse antibody (Santa Cruz, CA, USA), anti-Nrf2 antibody (Servicebio), anti-Keap1 rabbit pAb (Servicebio), anti-Lamin B1 rabbit pAb (Servicebio), and anti-β-actin antibody (Abcam, Cambridge, UK).

Techniques: Expressing, Infection