Journal: International Journal of Molecular Medicine

Article Title: HELLS inhibits autophagy-dependent ferroptosis in nasopharyngeal carcinoma by modulating the Nrf2/HO-1/GPX4 pathway

doi: 10.3892/ijmm.2026.5788

Figure Lengend Snippet: Effects of HELLS on ferroptosis. Levels of (A) GSH, (B) MDA and (C) Fe 2+ in the sh-HELLS, sh-NC, sh-HELLS + Era, sh-NC + Era, sh-HELLS + Era + Fer-1 and sh-NC + Era + Fer-1 groups. (D) Representative images illustrating immunofluorescence staining of ROS probes in the six groups. (E) ROS levels in the six groups. (F and G) Relative mRNA expression levels of (F) ACSL4 and (G) PTGS2 in the six groups. (H) Representative images of immunoblots of 4-HNE, ACSL4 and PTGS2. (I) Relative expression levels of 4-HNE in the six groups. (J and K) Relative protein expression levels of (J) ACSL4 and (K) PTGS2 in the six groups. * P<0.05, ** P<0.01, *** P<0.001 and **** P<0.0001. HELLS, lymphoid-specific helicase; GSH, glutathione; MDA, malondialdehyde; sh-, short hairpin; NC, negative control; Era, erastin; Fer-1, ferrostatin-1; ROS, reactive oxygen species; 4-HNE, 4-hydroxynonenal; ACSL4, acyl-CoA synthetase long-chain family member 4; PTGS2, prostaglandin-endoperoxide synthase 2.

Article Snippet: To modulate ferroptosis, cells transfected with sh-NC and sh-HELLS sequences were treated for 24 h with either 10 μ M of erastin (cat. no. HY-15763; MedChemExpress) or a combination of 10 μ M of Erastin and 2 μ M ferrostatin-1 (Fer-1; cat. no. HY-100579; MedChemExpress) ( , ).

Techniques: Immunofluorescence, Staining, Expressing, Western Blot, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: HELLS inhibits autophagy-dependent ferroptosis in nasopharyngeal carcinoma by modulating the Nrf2/HO-1/GPX4 pathway

doi: 10.3892/ijmm.2026.5788

Figure Lengend Snippet: Mechanisms underlying the effects of HELLS on autophagy-dependent ferroptosis. (A) Co-immunoprecipitation was performed in HK-1 cells with either HELLS antibody or normal rabbit IgG antibody, followed by immunoblot analysis using the indicated antibodies. (B) Relative viability of the sh-HELLS, sh-NC, sh-HELLS + ML334 and sh-NC + ML334 groups at 0, 24, 48 and 72 h. Levels of (C) GSH, MDA and (D) Fe 2+ in the four groups. (E-G) Relative mRNA expression levels of (E) GPX4, (F) HO-1 and (G) Nrf2 in the four groups. (H) Representative images of protein blots of GPX4, HO-1 and Nrf2. (I) Relative protein expression levels of GPX4 HO-1 and Nrf2 in the four groups. (J) Representative images of protein blots of Beclin1and LC3. (K) Relative protein expression levels of Beclin1and LC3 in the four groups. * P<0.05, ** P<0.01, *** P<0.001 and **** P<0.0001. HELLS, lymphoid-specific helicase; sh-, short hairpin; NC, negative control; GSH, glutathione; MDA, malondialdehyde; GPX4, glutathione peroxidase 4; HO-1, heme oxygenase-1; Nrf2, nuclear factor-erythroid 2-related factor 2.

Article Snippet: To investigate whether autophagy-dependent ferroptosis was regulated through the Nrf2/HO-1/GPX4 signaling pathway, cells were initially stimulated with erastin and subsequently treated with 1 μ M ML334 (cat. no. HY-110258; MedChemExpress) for 24 h.

Techniques: Immunoprecipitation, Western Blot, Expressing, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Hypothermic machine perfusion protects DCD graft liver from ischemia-reperfusion injury by enhancing macrophage efferocytosis via KLF2-NLRP3 signaling

doi: 10.3892/ijmm.2026.5756

Figure Lengend Snippet: KLF2 inhibits the NLRP3-mediated pyroptosis pathway during CS/Rep in ECs. (A) Heatmap and (B) volcano plot of differentially expressed genes between control and ov-KLF2 HUVECs after CS/Rep treatment. mRNA levels of (C) KLF2 and (D) NLRP3 in ov-control and ov-KLF2 group of HUVECs following CS/Rep treatment were quantified using reverse transcription-quantitative PCR. (E) Co-IP analysis of the interaction between KLF2 and NLRP3. (F) Microstructure of HUVECS. (G) Representative Western blot images for KLF2, NLRP3, GSDMD, Caspase-1 and IL-18 in CS/Rep HUVECs. Quantitative analysis of KLF2 (H), NLRP3 (I), GSDMD (J), Caspase-1 (K) and IL-18 (L) protein expression based on western blot results from HUVECS of ov-control and ov-KLF2 groups. Quantitative analysis of KLF2 (M), NLRP3 (N), GSDMD (O), Caspase-1 (P) and IL-18 (Q) protein expression based on western blot results from HUVECS of sh-control and sh-KLF2 groups. (n=3). * P<0.05, ** P<0.01, *** P<0.001. KLF2, Kruppel-like Factor 2; CS/Rep, cold storage/reperfusion; HUVEC, human umbilical vein endothelial cells; ov, overexpression; IP, immunoprecipitation; GSDMD, gasdermin D; sh, short hairpin;; FC, fold-change.

Article Snippet: The incubation period between transfection and subsequent treatment was 72 h. To verify whether NLRP3 inhibition rescues the phenotypes resulting from KLF2 deficiency, sh-KLF2-transfected cells were treated (37°C) with 10 μ M NLRP3 inhibitor MCC950 (cat. no. HY12815, MedChemExpress) for 24 h and subjected to CS/Rep as aforementioned.

Techniques: Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Co-Immunoprecipitation Assay, Western Blot, Expressing, Over Expression, Immunoprecipitation