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Fig. 3 CLA/CLNAs promote lysosomal degradation of GPX4. A, B Immunoblot of lysates from HT1080 cells treated with 10-CLA (200 µM) (A) or ESA (20 µM) (B) for indicated time using antibodies against GPX4 and β-actin. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13A, B. C HT1080 cells were treated with 10-CLA (200 µM) or ESA (20 µM) for 4 h, and relative mRNA level of GPX4 was measured by qRT-PCR analysis and shown as mean ± SD (n = 3). D–G Immunoblot of lysates from HT1080 cells pretreated with chloroquine (CQ: 20 µM) (D, E) or <t>MG132</t> (5 µM) (F, G) for 0.5 h, and treated with 10-CLA (200 µM) (D, F) or ESA (20 µM) (E, G) for indicated time, using antibodies against the indicated proteins. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13C–F.
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Fig. 3 CLA/CLNAs promote lysosomal degradation of GPX4. A, B Immunoblot of lysates from HT1080 cells treated with 10-CLA (200 µM) (A) or ESA (20 µM) (B) for indicated time using antibodies against GPX4 and β-actin. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13A, B. C HT1080 cells were treated with 10-CLA (200 µM) or ESA (20 µM) for 4 h, and relative mRNA level of GPX4 was measured by qRT-PCR analysis and shown as mean ± SD (n = 3). D–G Immunoblot of lysates from HT1080 cells pretreated with chloroquine (CQ: 20 µM) (D, E) or <t>MG132</t> (5 µM) (F, G) for 0.5 h, and treated with 10-CLA (200 µM) (D, F) or ESA (20 µM) (E, G) for indicated time, using antibodies against the indicated proteins. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13C–F.
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Fig. 3 CLA/CLNAs promote lysosomal degradation of GPX4. A, B Immunoblot of lysates from HT1080 cells treated with 10-CLA (200 µM) (A) or ESA (20 µM) (B) for indicated time using antibodies against GPX4 and β-actin. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13A, B. C HT1080 cells were treated with 10-CLA (200 µM) or ESA (20 µM) for 4 h, and relative mRNA level of GPX4 was measured by qRT-PCR analysis and shown as mean ± SD (n = 3). D–G Immunoblot of lysates from HT1080 cells pretreated with chloroquine (CQ: 20 µM) (D, E) or <t>MG132</t> (5 µM) (F, G) for 0.5 h, and treated with 10-CLA (200 µM) (D, F) or ESA (20 µM) (E, G) for indicated time, using antibodies against the indicated proteins. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13C–F.
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Fig. 3 CLA/CLNAs promote lysosomal degradation of GPX4. A, B Immunoblot of lysates from HT1080 cells treated with 10-CLA (200 µM) (A) or ESA (20 µM) (B) for indicated time using antibodies against GPX4 and β-actin. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13A, B. C HT1080 cells were treated with 10-CLA (200 µM) or ESA (20 µM) for 4 h, and relative mRNA level of GPX4 was measured by qRT-PCR analysis and shown as mean ± SD (n = 3). D–G Immunoblot of lysates from HT1080 cells pretreated with chloroquine (CQ: 20 µM) (D, E) or <t>MG132</t> (5 µM) (F, G) for 0.5 h, and treated with 10-CLA (200 µM) (D, F) or ESA (20 µM) (E, G) for indicated time, using antibodies against the indicated proteins. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13C–F.
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Image Search Results


Fig. 3 CLA/CLNAs promote lysosomal degradation of GPX4. A, B Immunoblot of lysates from HT1080 cells treated with 10-CLA (200 µM) (A) or ESA (20 µM) (B) for indicated time using antibodies against GPX4 and β-actin. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13A, B. C HT1080 cells were treated with 10-CLA (200 µM) or ESA (20 µM) for 4 h, and relative mRNA level of GPX4 was measured by qRT-PCR analysis and shown as mean ± SD (n = 3). D–G Immunoblot of lysates from HT1080 cells pretreated with chloroquine (CQ: 20 µM) (D, E) or MG132 (5 µM) (F, G) for 0.5 h, and treated with 10-CLA (200 µM) (D, F) or ESA (20 µM) (E, G) for indicated time, using antibodies against the indicated proteins. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13C–F.

Journal: Cell death & disease

Article Title: Conjugated fatty acids drive ferroptosis through chaperone-mediated autophagic degradation of GPX4 by targeting mitochondria.

doi: 10.1038/s41419-024-07237-w

Figure Lengend Snippet: Fig. 3 CLA/CLNAs promote lysosomal degradation of GPX4. A, B Immunoblot of lysates from HT1080 cells treated with 10-CLA (200 µM) (A) or ESA (20 µM) (B) for indicated time using antibodies against GPX4 and β-actin. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13A, B. C HT1080 cells were treated with 10-CLA (200 µM) or ESA (20 µM) for 4 h, and relative mRNA level of GPX4 was measured by qRT-PCR analysis and shown as mean ± SD (n = 3). D–G Immunoblot of lysates from HT1080 cells pretreated with chloroquine (CQ: 20 µM) (D, E) or MG132 (5 µM) (F, G) for 0.5 h, and treated with 10-CLA (200 µM) (D, F) or ESA (20 µM) (E, G) for indicated time, using antibodies against the indicated proteins. Images are cropped for clarity; full-length blots are presented in Supplementary Fig. 13C–F.

Article Snippet: All reagents were obtained from commercial suppliers: 1S,3R-RSL3 (#SML2234), Ferrostatin-1 (#SML0583) (Sigma, Burlington, MA, USA), mitoTEMPO (#sc-221945), Necrostatin-1 (#sc-200142), MG132 (#sc-201270) (Santa Cruz, Dallas, TX, USA), z-VAD-fmk (#3188-v) (Peptide Institute, Osaka, Japan), Necrostatin-1s (7-Cl-O-Nec-1, #S8641), Rucaparib (#S4948) (Selleck, Houston, TX, USA), Liperfluo (#L248), mitoPeDPP (#M466), FerroOrange (#F374) (Dojindo, Kumamoto, Japan), LipiRADICAL Green (#FDV-0042) (Funakoshi, Tokyo, Japan), MitoSOX (#M36008) (Invitrogen, Waltham, MA, USA), Erastin (#17754), Triacsin C (#10007448), PF-04620110 (iDGAT1, #16425), PF06424439 (iDGAT2, #17680), deferoxamine (DFO, #14595), Perhexiline (#16982), 2-cyano-3, 12-dioxooleana-1, 9(11)-dien-28-oate (CDDO, #81035) (Cayman, Ann Arbor, MI, USA), Chloroquine (#08660-04), Carbonyl Cyanide m-Chlorophenylhydrazone (CCCP, #07253-74) (Nacalai Tesque, Kyoto, Japan).

Techniques: Western Blot, Quantitative RT-PCR