Journal: Frontiers in Pharmacology

Article Title: Therapeutic Potential of Astragaloside IV Against Adriamycin-Induced Renal Damage in Rats via Ferroptosis

doi: 10.3389/fphar.2022.812594

Figure Lengend Snippet: ASIV alleviated iron accumulation in rats with ADR-induced renal damage (ADR disrupts iron metabolism, which leads to iron deposition in renal tissue). (A) Representative images of Perls’ Prussian blue staining and quantification of iron accumulation. (B) Representative Western blotting photographs. (C) Renal FPN1 expression. (D) The expression of DMT1. (E) The expression of renal TfR1. (F) Renal FTL expression. Data were expressed as mean ± SD, n = 6. * p < 0.05, ** p < 0.01 vs. CON group. # p < 0.05, ## p < 0.01 vs. ADR group.

Article Snippet: Immunoblotting was performed on the separated protein and probe with the following primary antibodies overnight at 4°C: TfR1 (1:2,000, Cat#abs131442, absin), FTL (1:10,000, Cat#3549-1, EPITMICS), DMT1 (1:2,000, Cat#abs112967, absin), FPN1 (1:3,000, Cat#GTX100573, absin), NOX2 (1:2,000, Cat#BA2811, BOSTER), NOX4 (1:1,000, Cat#GB11347, servicebio), GPx4 (1:2,000, Cat#ET1706-45, HUABIO), SLC7A11 (1:1,000, Cat#DF12509, Affinity), Nrf2 (1:1,000, Cat#M200-3, MBL), HO-1 (1:2,000, Cat#GB12104, servicebio), p-Pi3K (Cat#AF3241, Affinity), Pi3K (Cat#AF6241, Affinity), p-Akt (Cat#66444-1-Ig, Proteintech), Akt (Cat#60203-2-Ig, Proteintech), β-actin (1:1,000, Cat#CW0096S, CWBIO), and Lamin B1 (1:1,500, Cat#SI17-06, HUABIO).

Techniques: Staining, Western Blot, Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Hepcidin-to-Ferritin Ratio Is Decreased in Astrocytes With Extracellular Alpha-Synuclein and Iron Exposure

doi: 10.3389/fncel.2020.00047

Figure Lengend Snippet: Changes in iron metabolism-related proteins in α-syn-treated primary cultured astrocytes. Proteins levels of DMT1, FPN1, IRP1 and IRP2 were unchanged (A) . Quantitative analysis of the proteins was shown in (B) . Data are presented as the ratio of DMT1, FPN1, IRP1 and IRP2 protein levels to β-actin. Each bar represents the mean ± SEM of three independent experiments.

Article Snippet: Divalent metal transporter 1 (DMT1) antibody was from OriGene (Rockville, MD, USA), antibodies against ferritin, iron regulatory protein (IRP) 1 and IRP2 antibody were from Abcam (Cambridge, UK).

Techniques: Cell Culture

Journal: Cancers

Article Title: Ferroptosis Inducers Kill Mesenchymal Stem Cells Affected by Neuroblastoma

doi: 10.3390/cancers15041301

Figure Lengend Snippet: RNA-seq was performed and anti-ferroptosis pathways were selected. ( A , B ) The pathways linked with ferroptosis were selected. ( C , D ) Western blotting was used to analyze protein levels of NRF2, SLC7A11 and GPX4. ( E ) GPx activity was detected in BMSCs and NB-BMSCs. A one-way analysis of variance was used. ns not significant; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Immunoblotting was performed overnight at 4 °C using primary antibodies against SLC7A11 (26864-1-AP, Proteintech, Rosemont, IL, USA), NFS1 (15370-1-AP, Proteintech), GPX4 (ab125066, abcam, Cambridge, UK), Ferritin heavy chain (FTH, ab75972, abcam), Ferritin light chain (FTL, ab75973, abcam), SLC40A1 (FPN, ab239583, abcam), Mitochondrial ferritin (MTFT, ab124889, abcam), Transferrin receptor (TFR, ab214039, abcam), SLC11A2 (divalent metal transporter 1, DMT1, 20507-1-AP, Proteintech), NRF2 (16396-1-AP, Proteintech), and GAPDH (60004-1-Ig, Proteintech).

Techniques: RNA Sequencing Assay, Western Blot, Activity Assay

Journal: Cancers

Article Title: Ferroptosis Inducers Kill Mesenchymal Stem Cells Affected by Neuroblastoma

doi: 10.3390/cancers15041301

Figure Lengend Snippet: Immunofluorescence indicated decreased anti-ferroptosis capacity of BMSCs in patients with BM metastasis. ( A – F ) CD90 and CD105 were used to mark BMSCs; SLC7A11, GPX4, and NFS1 were detected in BM of NB patients with or without BM metastasis. Scale bar: 50 μm. Student’s t -test was used in the analysis. *** p < 0.001.

Article Snippet: Immunoblotting was performed overnight at 4 °C using primary antibodies against SLC7A11 (26864-1-AP, Proteintech, Rosemont, IL, USA), NFS1 (15370-1-AP, Proteintech), GPX4 (ab125066, abcam, Cambridge, UK), Ferritin heavy chain (FTH, ab75972, abcam), Ferritin light chain (FTL, ab75973, abcam), SLC40A1 (FPN, ab239583, abcam), Mitochondrial ferritin (MTFT, ab124889, abcam), Transferrin receptor (TFR, ab214039, abcam), SLC11A2 (divalent metal transporter 1, DMT1, 20507-1-AP, Proteintech), NRF2 (16396-1-AP, Proteintech), and GAPDH (60004-1-Ig, Proteintech).

Techniques: Immunofluorescence