Journal: Antioxidants (Basel, Switzerland)

Article Title: OTUB1-SLC7A11 Axis Mediates 4-Octyl Itaconate Protection Against Acetaminophen-Induced Ferroptotic Liver Injury.

doi: 10.3390/antiox14060698

Figure Lengend Snippet: Figure 3. 4-OI mitigates APAP-induced dysregulation of iron metabolism. (A) Pathways associated with genes/proteins interacting with differential metabolites were analyzed using the R package CePa. logP (x-axis): negative log of the enrichment p-value for drug bias-related metabolites. foldP (y-axis): ratio of enrichment p-values for efficacy-related vs. bias-related metabolites. The red circle highlights the ferroptosis pathway, which is identified as the most prominent among all analyzed pathways. (B) Hepatic iron content (n = 3). (C–I) Representative Western blot images and quantification of transferrin, TFR1, FTH1, FTL1, FPN1, and hepcidin protein levels in mouse liver tissues (n = 3). (J–L) A RT-qPCR assay determined the mRNA expression of FTH1, FTL1, and FPN1 (n = 6). (M) Representative immunohistochemical staining images showing FTH1, FTL1, and FPN1 expression. Scale bar: 100 µm, n = 3. GAPDH was used as internal loading control. * p < 0.05, ** p < 0.01, *** p < 0.001; ns: not significant.

Article Snippet: Primary antibodies for SLC7A11, OTUB1, CD44, ubiquitin, GPX4, Nrf2, transferrin, TFR1, FTH1, FTL1, FPN1, hepcidin, and GAPDH were obtained from the following sources: SLC7A11 and OTUB1 antibodies from Abcam (Cambridge, UK); hepcidin antibody from Affinity (Changzhou, China); CD44, ubiquitin, GPX4, Nrf2, transferrin, TFR1, FTH1, FTL1, FPN1, and GAPDH antibodies from Proteintech (Wuhan, China).

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Immunohistochemical staining, Staining, Control

Journal:

Article Title: Phospholipase D2 Is Localized to the Rims of the Golgi Apparatus in Mammalian Cells

doi: 10.1091/mbc.02-04-0059

Figure Lengend Snippet: PLD2 localization with different organelles in rat NRK cells. Cells were prepared for immunofluorescence microscopy and incubated with rabbit PLD2 antibody PLD2-27 (A, D, G, and J) and costained with monoclonal antibodies to the ER marker BiP (B); the transferrin receptor (E), a marker of early endosomes and the plasma membrane; and lgp120 (H), a late endosome/lysosomal marker. (K) Cells were stained with caveolin-1, a marker for caveoli; the arrow corresponds to caveolin-1 localized to the plasma membrane. Images were merged to determine overlap between PLD2 (red) and the respective marker proteins (green; C, F, I, and L). Areas of maximal overlap are yellow. (L) Inset, enlargement of the region of overlap between PLD2 and caveolin-1 in the perinuclear Golgi apparatus. The arrow indicates areas where PLD2 puncta (red) either overlap completely with caveolin-1 (green) or are directly adjacent. Images are from projected Z-series. Bars, 10 μm.

Article Snippet: Purified monoclonal anti-rat transferrin receptor antibodies were purchased from Cedarlane Laboratories (Hornby, Ontario, Canada).

Techniques: Immunofluorescence, Microscopy, Incubation, Marker, Staining

Journal: Redox Biology

Article Title: Differentiation of intestinal stem cells toward goblet cells under systemic iron overload stress are associated with inhibition of Notch signaling pathway and ferroptosis

doi: 10.1016/j.redox.2024.103160

Figure Lengend Snippet: List of antibodies used in Western blot analysis and immunofluorescence microscopy.

Article Snippet: Anti-TfR , Bioss , bs-0988R , WB.

Techniques: Western Blot, Immunofluorescence, Microscopy