Journal: bioRxiv

Article Title: Iron addicted colorectal cancers exploit Heme-Complex II axis to resist oxidative cell death

doi: 10.1101/2025.11.05.686813

Figure Lengend Snippet: A) Western blot images showing protein expression of UROD and Vinculin in HCT116 and SW480 cells stable for empty tet-pLKO-puro (Scr) or UROD targeting shRNA (shRNA1 and shRNA2) 24h, 48h and 72h post dox treatment (250ng/mL). B) Representative crystal violet staining image in cells from b) 14 days post FAC treatment (0.25, 0.5mM) with or without dox (250ng/mL). C) Measurement of relative growth by quantification of relative colony count from (B). D) Western blot images showing protein expression of FECH and Vinculin in HCT116 and SW480 cells stable for empty tet-pLKO-puro (Scr) or FECH targeting shRNA (shRNA1 and shRNA2) 24h, 48h and 72h post dox treatment (250ng/mL). E) Representative crystal violet staining image in cells from e) 14 days post FAC treatment (0.25, 0.5mM) with or without dox (250ng/mL) or heme (10μM). F) Measurement of relative growth by quantification of relative colony count from (E). G) Representative crystal violet staining image in human CRC cells from 14 days post FAC treatment with or without SA (250μM) and Heme (10μM). All data are mean ± SEM. One-way ANOVA with Tukey’s multiple comparisons test for (C), and (F). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After lysis solubilized proteins were resolved on 12% SDS-polyacrylamide gels and transferred to PVDF membrane, blocked with 5% milk in TBST and probed overnight with indicated primary antibodies diluted at 1:1000 in the blocking buffer for GPX4 (Proteintech, 67763-1-Ig), FTH1 (Cell Signaling,3998S), UROD (Santa Cruz, SC-365297), FECH (Proteintech, 14466-1-AP), SDHA (Cell Signaling, 11998), SDHB (Proteintech, 10620-1-AP), SDHC (Proteintech, 14575-AP), β-actin (Proteintech 66009-1-Ig).

Techniques: Western Blot, Expressing, shRNA, Staining

Journal: International Journal of Molecular Sciences

Article Title: Relationship of Iron Metabolism and Short-Term Cuprizone Treatment of C57BL/6 Mice

doi: 10.3390/ijms20092257

Figure Lengend Snippet: mRNA and protein analyses of NFS1, frataxin (FRX), and ferrochelatase (Fc) genes in CZ treated groups. Real Time PCR was performed with SYBR green protocol using gene specific primers. The relative expression of the controls was regarded as 1. Western blot analyses were done from protein extracts of the liver tissue samples. The same amount of protein (15 µg) from each sample was loaded onto 10% SDS-PAGE and transferred to nitrocellulose membranes and probed with NFS1 or Fc polyclonal rabbit antibodies. β-actin was used as loading control. ( A ) Relative mRNA expression values of NFS1, FRX, and Fc in CC. ( B ) Relative mRNA levels of NFS1, FRX, and Fc in the liver. ( C ) Protein expression levels of NFS1 and Fc in the liver. The Western blot experiments were repeated at least three times in each group. The bars represent mean values and error bars represent standard errors of the mean (SEM) for three independent measurements. For statistical analyses, Student’s t -test and two-sample t -tests were used. The asterisk marks p < 0.05 compared to non-treated animals, while the cross indicates a difference p < 0.05 between the treated groups, n = 6 in each group. Optical density analyses can be seen in .

Article Snippet: The same amount of protein from each sample was loaded onto 12% or 14% SDS-PAGE and transferred to nitrocellulose membranes (BioTrace NT, Pall Life Sciences, Port Washington, NY) and probed with rabbit polyclonal antibodies produced against mouse ferrochelatase (Fc 1:1000; Novus Biologicals), mitochondrial ferritin (FTMT 1:1000; Thermo-Fischer Scientific Inc.), ferritin heavy chain (FTH 1:1000; Cell Signaling Technology Europe, Leiden, The Netherlands), NFS1 (1:1000; Novus Biologicals), transferrin receptor-1 (TfR1 1:1000; Thermo-Fischer Scientific Inc.) and alpha-1 antitrypsin (A1AT 1:1000; Abcam, Cambridge, UK). β-actin (1:2000; Sigma-Aldrich Kft.) was used as loading control.

Techniques: Real-time Polymerase Chain Reaction, SYBR Green Assay, Expressing, Western Blot, SDS Page, Control