ch55  (TargetMol)

Journal: Acta Pharmaceutica Sinica. B

Article Title: Vitamin A and its analogues modulate MUFAs metabolism to improve ferroptosis and aging by direct targeting of ACSL3

doi: 10.1016/j.apsb.2025.11.004

Figure Lengend Snippet: Vitamin A and its metabolites inhibit ferroptosis independent of antioxidant capacity or RAR/RXR activation. (A) Illustration of the structures of VA and ATRA. (B) Measurement of the EC 50 values of VA and ATRA towards RSL3 (0.5 μmol/L) or Erastin (10 μmol/L) induced ferroptosis in HT-1080 cells. Cells were treated with drugs for 24 h. (C) RAL suppressed RSL3 and Erastin-induced ferroptosis in HT-1080 cells. (D) Viability of HT-1080 GPX4 -KO cells treated with the indicated compounds for 24 h after withdrawal of Fer-1, which is essential for the survival of GPX4 -KO cells. (E) The antioxidative capacity of VA (50 μmol/L), ATRA (50 μmol/L) and RAL (50 μmol/L) was analyzed by the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. Trolox (50 μmol/L) was used as a positive control. (F) The effect of VA, ATRA and RAL on the autoxidation of C11 BODIPY 581/591 initiated by AAPH in EggPC liposomes was measured by FENIX assay. Fer-1 was used as a positive control. (G) Protein expression levels of GPX4, SLC7A11, ACSL4, SCD1, FSP1, HO-1 and NRF2 in HT-1080 cells were analyzed by Western blot. Cells were treated with VA (1 μmol/L) or ATRA (10 μmol/L) in the presence or absence of RSL3 (0.25 μmol/L) for 2.5 h or Erastin (10 μmol/L) for 8 h. (H) Protein expression levels of IRP1, TFRC and FTH1 in HT-1080 cells were analyzed by Western blot. Cells were treated with VA (1 μmol/L) or ATRA (10 μmol/L) in the presence or absence of RSL3 (0.25 μmol/L) for 2.5 h or Erastin (10 μmol/L) for 8 h. (I, J) Cell viability of HT-1080 cells after co-treatment with RSL3 and VA or ATRA in the presence of pan-RAR antagonist AGN193109 (20 μmol/L) (I) or pan-RXR antagonist HX531 (20 μmol/L) (J). (K) The protective effects of VA or ATRA on RSL3-induced ferroptosis in RXRA and RXRB knockout HT-1080 cells. (L) Viability of HT-1080 cells treated with different doses of RAR agonists Ch55 and Adapalene (ADA) under RSL3 (0.5 μmol/L) and Erastin (10 μmol/L)-induced ferroptosis for 24 h. Data shown represent the mean ± SD ( n = 3). ∗∗ P < 0.01, and ∗∗∗ P < 0.001, indicating significant differences between groups. ns means no significance.

Article Snippet: All- trans -retinal (T5256), Fenretinide (T1872), Acitretin (T1330), AGN193109 (TQ0097), HX531( T22843 ), Ch55 ( T14946 ), Adapalene (T1093) and A939572 (T4515) were purchased from TargetMOI (Shanghai, China).

Techniques: Activation Assay, Positive Control, Liposomes, Expressing, Western Blot, Knock-Out

Journal: Communications Biology

Article Title: Retinoic acid receptor alpha inhibits ferroptosis by promoting thioredoxin and protein phosphatase 1F in lung adenocarcinoma

doi: 10.1038/s42003-024-06452-7

Figure Lengend Snippet: a CCK8 assays to detect the cell viability of A549 and PC9 cells treated with RSL3 (2 μM), RSL3 (2 μM) plus DFO (100 μM) or fer-1 (20 μM) for 24 h after incubating with DMSO, Ch55 (5 μM), AM580 (5 μM) ATRA (20 μM) or AGN193109 (5 μM) for 24 h. ( n = 4 biologically independent experiments, Student t -test). b – e MDA level ( b ) lipid peroxidation ( c ), mitochondrial membrane potential (MMP) by TMRE ( d ), and labile iron pool (LIP) by CA-AM ( e ) were accessed in A549 and PC9 cells treated with DMSO or RSL3 (1 μM) for 24 h after incubating with DMSO, Ch55 (5 μM), AM580 (5 μM) ATRA (20 μM) or AGN193109 (5 μM) for 24 h. ( n = 3 biologically independent experiments, Student t -test). f Light microscopy images showed the degrees of “ballooning phenotype” in A549 and PC9 cells treated with DMSO or RSL3 (1 μM) for 24 h after incubating with DMSO, Ch55 (5 μM), AM580 (5 μM) ATRA (20 μM) or AGN193109 (5 μM) for 24 h. Scale bar:50 μm. The zoomed-in figures, scale bar: 250 μm. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: The RSL3, Imidazole Ketone Erastin (IKE), Erastin, Ferric ammonium citrate (FAC), RARA inhibitor AGN193109, RARA activator Ch55 and AM580, PAK inhibitor FRAX486, cisplatin (CDDP), and pemetrexed were acquired from Topscience (Shanghai, China) and were dissolved in dimethyl sulfoxide (DMSO).

Techniques: Membrane, Light Microscopy

Journal: Communications Biology

Article Title: Retinoic acid receptor alpha inhibits ferroptosis by promoting thioredoxin and protein phosphatase 1F in lung adenocarcinoma

doi: 10.1038/s42003-024-06452-7

Figure Lengend Snippet: a qRT-PCR and WB assays confirmed the mRNA and protein expression level of RARA in A549 and PC9 cells after transfection with NC (control shRNA), RARA-SH1, or RARA-SH2 lentivirus. ( n = 3 biologically independent experiments, Student t -test) ( b ) CCK8 assays to detect the cell viability of NC, RARA-SH1, or RARA-SH2 groups in A549 and PC9 cells treated with a gradient dose of RSL3 or IKE for 24 h. ( n = 4 biologically independent experiments, two-way ANOVA) ( c ) CCK8 assays to detect the cell viability of NC, RARA-SH1, or RARA-SH2 groups in A549 and PC9 cells treated with DMSO or RSL3 (2 μM) for 24 h after incubating with DMSO, Ch55 (5 μM), AM580 (5 μM) ATRA (20 μM) or AGN193109 (5 μM) for 24 h. ( n = 4 biologically independent experiments, Student t -test). d – g MDA level ( d ), lipid peroxidation ( e ), MMP by TMRE ( g ) and LIP by CA-AM ( g ) were accessed in NC, RARA-SH1 groups of A549 and PC9 cells treated with DMSO or RSL3 (1 μM) for 24 h after incubating with DMSO, Ch55 (5 μM), AM580 (5 μM) ATRA (20 μM) or AGN193109 (5 μM) for 24 h. ( n = 3 biologically independent experiments, Student t -test) ns, not significant * p < 0.05; ** p < 0.01; *** p < 0.001;**** p < 0.0001.

Article Snippet: The RSL3, Imidazole Ketone Erastin (IKE), Erastin, Ferric ammonium citrate (FAC), RARA inhibitor AGN193109, RARA activator Ch55 and AM580, PAK inhibitor FRAX486, cisplatin (CDDP), and pemetrexed were acquired from Topscience (Shanghai, China) and were dissolved in dimethyl sulfoxide (DMSO).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Control, shRNA