fe 2+ content  (Beijing Solarbio Science)

Journal: Frontiers in Pharmacology

Article Title: Targeted ferroptosis of myofibroblasts by tubeimoside I attenuates hypertrophic scar formation

doi: 10.3389/fphar.2025.1654108

Figure Lengend Snippet: TBMS1 caused HSF ferroptosis. (A) The volcano plot displays significantly differentially-expressed genes (DEGs) in TBMS1-treated HSFs. (B) GSEA of DEGs in TBMS1-treated HSFs. Pathways related to PI3K/AKT pathway and ferroptosis are presented. (C) The heatmap shows typical TBMS1-regulated ferroptosis genes in HSFs. (D) The expression of the mRNAs of DEGs associated with ferroptosis in HSFs treated with vehicle or TBMS1 was estimated by qRT-PCR. (E) HSFs were treated with TBMS1 or TBMS1 combined with 1 µM ferrostatin-1, 10 µM Z-VAD-FMK, or 1 μM necrosulfonamide for 48 h before the measurement of cell viability (n = 3 separate assays). (F) ROS and MDA levels, intracellular Fe 2+ contents, and the GSH/GSSG ratio in vehicle-treated or TBMS1-treated HSFs (n = 3 separate assays). (G) ROS and MDA levels, intracellular Fe 2+ contents, and GSH/GSSG ratio of HSFs after treatment with TBMS1 or TBMS1 combined with ferrostatin-1 (1 µM) (n = 3 separate assays). (H) Western blotting illustrated TFR1 and DMT1 levels within HSFs after vehicle or TBMS1 treatment. (I) the Fe 2+ /Fe 3+ ratio in vehicle-treated or TBMS1-treated HSFs (n = 3 separate assays). (J) the Fe 2+ /Fe 3+ ratio of HSFs after treatment with TBMS1 or TBMS1 combined with ferrostatin-1 (1 µM) (n = 3 separate assays). The data suggest mean ± SD. Experiments (E–J) were examined via one-way ANOVA based on Tukey’s multiple comparison test, whereas assay (D) was performed via an unpaired two-tailed Student’s t-test; *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: After inoculation (1.2 × 10 6 /well) in six-well plates, the cells were subjected to relevant treatments for 48 h. Intracellular Fe 2+ contents were analyzed with the ferrous iron colorimetric assay kit (E-BC-K881-M, Elabscience, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Comparison, Two Tailed Test

Journal: Redox Biology

Article Title: The role and mechanism of the cGAS–STING pathway-mediated ROS in apoptosis and ferroptosis induced by manganese exposure

doi: 10.1016/j.redox.2025.103761

Figure Lengend Snippet: Effects of MnCl 2 and Erastin on ferroptosis in BV2 cells. (A) Heatmap analysis of differentially expressed ferroptosis related genes screened by RNA-seq. (B, C) Detection and relative quantitative analysis of mitochondrial membrane potential in BV2 cells by flow cytometry ( n = 3). (D) Fe 2+ content in BV2 cells ( n = 3). (E–J) Western blot was conducted to detect the expression of GPX4, SLC7A11, FSP1, DHODH, ACSL4 and quantitative analysis ( n = 3). (K) Fe 2+ content in BV2 cells treated with Erastin combined with MnCl 2 ( n = 3). (L–Q) Western blot was conducted to detect the expression of GPX4, SLC7A11, FSP1, DHODH and ACSL4 in BV2 cells treated with Erastin combined with MnCl 2 and quantitative analysis ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The Fe 2+ content was calculated according to the instructions (Elabscience, Wuhan, China), and the experiment was repeated three times.

Techniques: RNA Sequencing, Membrane, Flow Cytometry, Western Blot, Expressing

Journal: Redox Biology

Article Title: The role and mechanism of the cGAS–STING pathway-mediated ROS in apoptosis and ferroptosis induced by manganese exposure

doi: 10.1016/j.redox.2025.103761

Figure Lengend Snippet: Mn exposure induces mitochondrial ultrastructural disruption, cGAS–STING pathway activation, and coordinated dysregulation of oxidative stress, apoptosis, and ferroptosis in mouse hippocampus. (A) Transmission electron microscopy was used to observe the effect of Mn exposure on the ultrastructure of mitochondria in the hippocampus of mice. Yellow arrows indicate cell membranes and mitochondria. Scales are 2 μm (6,000✕) and 500 nm (30,000✕). (B–D) Western blot was conducted to detect the expression of TBK1 and IRF3 in mouse brain tissue and quantitative analysis ( n = 3). (E–G) Detection of GSH-Px, SOD activity and MDA content in mouse brain tissue ( n = 3). (H–J) Western blot was conducted to detect the expression of Bax and Cytochrome C in mouse brain tissue and quantitative analysis ( n = 3). (K) Fe 2+ content in mouse brain tissue ( n = 3). (L) Immunofluorescence detection of ACSL4 expression in mouse brain tissue (40 × ). (M − P) Western blot was conducted to detect the expression of GPX4、ACSL4 and DHODH in mouse brain tissue and quantitative analysis ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The Fe 2+ content was calculated according to the instructions (Elabscience, Wuhan, China), and the experiment was repeated three times.

Techniques: Disruption, Activation Assay, Transmission Assay, Electron Microscopy, Western Blot, Expressing, Activity Assay, Immunofluorescence

Journal: Redox Biology

Article Title: The role and mechanism of the cGAS–STING pathway-mediated ROS in apoptosis and ferroptosis induced by manganese exposure

doi: 10.1016/j.redox.2025.103761

Figure Lengend Snippet: Suppression of the cGAS–STING pathway mitigates MnCl 2 -induced ferroptosis. (A, I) Heat map analysis of differentially expressed genes of ferroptosis within BV2 cGAS−/− and BV2 STING−/− screened by RNA-seq. (B, J) Fe 2+ content in BV2 cGAS−/− and BV2 STING−/− cell lines ( n = 3). (C–H, K–P) Western blot was conducted to detect the expression of GPX4, SLC7A11, FSP1, DHODH and ACSL4 in BV2 cGAS−/− and BV2 STING−/− and quantitative analysis ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The Fe 2+ content was calculated according to the instructions (Elabscience, Wuhan, China), and the experiment was repeated three times.

Techniques: RNA Sequencing, Western Blot, Expressing

Journal: Redox Biology

Article Title: The role and mechanism of the cGAS–STING pathway-mediated ROS in apoptosis and ferroptosis induced by manganese exposure

doi: 10.1016/j.redox.2025.103761

Figure Lengend Snippet: Role of cGAS–STING pathway-mediated ROS in MnCl 2 -induced apoptosis and ferroptosis in BV2 cells. (A) The ROS level of BV2 cells treated with NAC combined with MnCl 2 was detected by biochemical kit ( n = 5). (B, C) ROS level and fluorescence intensity analysis of BV2 cells treated with NAC combined with MnCl 2 by flow cytometry ( n = 3). (D, E) Apoptosis level and relative quantitative analysis of BV2 cells detected by flow cytometry ( n = 3). (F–H) Western blot was conducted to detect the expression of Bcl-2 and Cytochrome C in BV2 cells treated with NAC combined with MnCl 2 and quantitative analysis ( n = 3). (I) Fe 2+ content in BV2 cells treated with NAC combined with MnCl 2 ( n = 3). (J–M) Western blot was conducted to detect the expression of GPX4, SLC7A11 and ACSL4 in BV2 cells treated with NAC combined with MnCl 2 and quantitative analysis ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The Fe 2+ content was calculated according to the instructions (Elabscience, Wuhan, China), and the experiment was repeated three times.

Techniques: Fluorescence, Flow Cytometry, Western Blot, Expressing

Journal: Journal of Cell Communication and Signaling

Article Title: ELAVL1 promotes ferroptosis via the TRIM21/HOXD8 axis to inhibit osteogenic differentiation in congenital pseudoarticular tibia‐derived mesenchymal stem cells

doi: 10.1002/ccs3.70016

Figure Lengend Snippet: Knockdown of TRIM21 inhibited ferroptosis to promote osteogenic differentiation of CPT mesenchymal stem cells (MSCs) were derived from normal iliac periosteum and CPT patient periosteum, respectively ( n = 3). (A) Western blot was performed to detect TRIM21 protein expression; (B) expression and distribution of TRIM21 wee analyzed using IF staining (scale bar = 100 μm). CPT MSCs were infected with lenti‐sh‐TRIM21 or lenti‐sh‐NC and induced osteogenic differentiation. (C) TRIM21 expression was tested using the western blot; (D) the treated CPT MSCs were subjected to alkaline phosphatase staining (scale bar = 100 μm); (E) mineralization of the cellular matrix was examined through ARS staining (scale bar = 100 μm); (F) osteogenesis‐related marker genes RUNX2, OPN, and OCN were examined using the western blot. Erastin (10 μM) was added to the above groups of cells to activate ferroptosis in MSCs and (G and H) Fe 2+ content and lipid reactive oxygen species content in MSCs were determined using the kit; (I) GPX4 and SLC7A11 expression in MSCs were checked using the western blot. n = 3. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Upon successful lentiviral infection, MSCs (1 × 10 4 cells) were treated with erastin (10 μM) for 48 h. Subsequently, experiments were performed following Fe 2+ content assay kit (BC5415, Solarbio, Beijing, China) instructions.

Techniques: Knockdown, Derivative Assay, Western Blot, Expressing, Staining, Infection, Marker

Journal: Journal of Cell Communication and Signaling

Article Title: ELAVL1 promotes ferroptosis via the TRIM21/HOXD8 axis to inhibit osteogenic differentiation in congenital pseudoarticular tibia‐derived mesenchymal stem cells

doi: 10.1002/ccs3.70016

Figure Lengend Snippet: Knockdown of ELAVL1 suppressed ferroptosis to promote osteogenic differentiation of CPT mesenchymal stem cells (MSCs). (A) ELAVL1 expression in normal MSCs and CPT MSCs were assessed using the western blot. (B) Expression and localization of ELAVL1 were analyzed in normal MSCs and CPT MSCs using the IF staining (scale bar = 100 μm). CPT MSCs were infected with lenti‐sh‐ELAVL1 or lenti‐sh‐NC and induced osteogenic differentiation. (C) Western blot detection of ELAVL1 expression; (D) alkaline phosphatase (ALP) staining assay experiments using the ALP staining kit (scale bar = 100 μm); (E) cellular matrix mineralization was examined using the ARS staining kit (scale bar = 100 μm); (F) RUNX2, OPN, and OCN expression were assessed using the western blot. (G and H) Fe 2+ content and lipid reactive oxygen species level in erastin‐treated MSCs were measured using the kit; (I) Western blot was applied to analyze ferroptosis‐related factors and GPX4 and SLC7A11 expression in erastin‐treated MSCs. n = 3. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Upon successful lentiviral infection, MSCs (1 × 10 4 cells) were treated with erastin (10 μM) for 48 h. Subsequently, experiments were performed following Fe 2+ content assay kit (BC5415, Solarbio, Beijing, China) instructions.

Techniques: Knockdown, Expressing, Western Blot, Staining, Infection

Journal: Journal of Cell Communication and Signaling

Article Title: ELAVL1 promotes ferroptosis via the TRIM21/HOXD8 axis to inhibit osteogenic differentiation in congenital pseudoarticular tibia‐derived mesenchymal stem cells

doi: 10.1002/ccs3.70016

Figure Lengend Snippet: ELAVL1 promoted ferroptosis by regulating the TRIM21/HOXD8 axis to inhibit osteogenic differentiation of CPT mesenchymal stem cells (MSCs). Lenti‐sh‐ELAVL1 or/and lenti‐TRIM21 were utilized to infect CPT MSCs. (A) The expression of ELAVL1, TRIM21, and HOXD8 was assayed with the western blot method. (B) Alkaline phosphatase staining assays were performed using kits (scale bar = 100 μm). (C) ARS staining was used to examine mineralized nodules (scale bar = 100 μm). (D) Osteogenesis‐related marker genes RUNX2, OPN, and OCN expression were measured using the western blot. (E and F) Fe 2+ content and lipid reactive oxygen species content of erastin‐treated CPT MSCs were assayed using the kit. (G) Western blot was applied to examine GPX4 and SLC7A11 expression in erastin‐treated CPT MSCs. n = 3. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Upon successful lentiviral infection, MSCs (1 × 10 4 cells) were treated with erastin (10 μM) for 48 h. Subsequently, experiments were performed following Fe 2+ content assay kit (BC5415, Solarbio, Beijing, China) instructions.

Techniques: Expressing, Western Blot, Staining, Marker