Journal: Journal of the American Chemical Society

Article Title: PROTAC-Mediated Degradation of mHTT Aggregates Attenuates Neurotoxicity in Cellular and R6/2 Mouse Models of Huntington’s Disease

doi: 10.1021/jacs.5c14078

Figure Lengend Snippet: PROTAC 2′ administration by osmotic pump decreases mHTT aggregates and inflammation in the R6/2 mouse brain. Representative images of immunofluorescence staining (anti-EM48) of the cortical (A) and striatal (B) regions of the R6/2 mouse brain with or without PROTAC 2′ treatment. Quantification of mHTT puncta amounts (C) and size (D) in the brain regions in panels A and B. Representative images of immunofluorescence staining (anti-Iba1) of WT (E) and R6/2 (F) mouse brains with or without PROTAC 2′ treatment. (G) Quantification of inflammation level in panels E and F. Representative images of immunofluorescence staining (anti-DARPP32) of WT (H) and R6/2 (I) mouse brains with or without PROTAC 2′ treatment. (J) Quantification of DARPP32 level in panel H and I. All the statistical results were quantified by ImageJ and shown as mean ± SD. The region for statistical analysis was arbitrarily selected (mouse number = 3, each dot represents an independent mouse brain; the technical replicates are shown by the violin plot, the selected region for analysis = 6). Panel C was analyzed by a two-tailed Welch’s t -test due to the unequal variance (** P < 0.01). Panel D was analyzed by a two-tailed unpaired t -test. Panels G and J were analyzed by one-way ANOVA with Tukey’s posthoc test (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: For immunofluorescence staining, sections were incubated overnight at 4 °C with EM48 (1:250, Merck Millipore, MAB5374), IBA1 (1:250, Proteintech, 10904-1-ap), DARPP-32 (1:50, Cell Signaling, #2302), CD68 (1:100, Abcam, ab31630), and Galectin-3 B2C10 (1:200, Invitrogen, MA1-40229) primary antibodies.

Techniques: Immunofluorescence, Staining, Two Tailed Test

Journal: Biochemistry and Biophysics Reports

Article Title: Alantolactone triggers oxeiptosis in human ovarian cancer cells via Nrf2 signaling pathway

doi: 10.1016/j.bbrep.2023.101537

Figure Lengend Snippet: Western blotting shows KEAP1-PGAM5-AIFM1 pathway protein level regulation by ALT. SKOV-3 cells were treated with different concentrations (0, 16, 32 and 64 μM) of ALT for 24 h. Western blot images ( A ) analysis showed that Ala decreased protein levels of B. Nrf2 and C. P-AIFM1(Ser-116), phospho-specific. ALT increased protein levels of D. KEAP1, E. AIFM1and F. PGAM5. Data expressed as mean ± SD of three independent experiments. The significance was assessed with a one-way ANOVA test. For p-value each symbol is shown significance to control group. (* p < 0.05, ** p < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: The antibody against AIFM1 (E-AB-10123) was purchased from Elabscience biotechnology.

Techniques: Western Blot, Control

Journal: Biochemistry and Biophysics Reports

Article Title: Alantolactone triggers oxeiptosis in human ovarian cancer cells via Nrf2 signaling pathway

doi: 10.1016/j.bbrep.2023.101537

Figure Lengend Snippet: ALT inhibits Nrf2 transcription and elevates ROS production.ALT suppresses the transcription factor Nrf2 expression in SKOV3. Nrf2 stimulates the transcription of numerous cytoprotective genes such as glutathione synthetase and glutathione peroxidase. Downregulation of Nrf2 leads to a decrease in GSH/GPX mRNA levels which enhances ROS. Elevated ROS concentration lead to the dissociation of PGAM5 from KEAP1 and its translocation into the mitochondria. PGAM5 can dephosphorylates AIFM1 at S116. Dephosphorylated AIFM1is transferred to the nucleous and induces oxieptosis cell death pathway.

Article Snippet: The antibody against AIFM1 (E-AB-10123) was purchased from Elabscience biotechnology.

Techniques: Expressing, Concentration Assay, Translocation Assay