Journal: Scientific reports

Article Title: Orthogonal validation of PROTAC mediated degradation of the integral membrane proteins EGFR and c-MET.

doi: 10.1038/s41598-024-84217-2

Figure Lengend Snippet: Fig. 4. Degradation profile of wildtype EGFR in A549 cells. Cells were pre-treated as described in (Fig. 1) with EGF as RTK ligand and either PROTAC3 (A,C,E,G,I) or Gefitinib (B,D,F,H,J) was added for 24 h. Cells were then analysed by immunoblotting (A,B) with quantification (C,D), immunofluorescence for the whole cell (E,F) or plasma membrane only (G,H) or flow cytometry (I,J). Graphs show the mean SEM of at least 3 biological replicates. The dotted line indicates a value of 0.8. All details regarding statistical analyses and p-values are provided in the (Supplementary Table 3).

Article Snippet: For starvation experiments, cells were washed twice in PBS and grown in respective growth media without FBS for 6–8 h. Subsequently media were exchanged for full growth media and cells were treated with SJF8240 (Tocris, 7266), Foretinib (Stratech, ORB322222), PROTAC3 (Tocris, 7258) or Gefitinib (AstraZeneca).

Techniques: Western Blot, Immunofluorescence, Clinical Proteomics, Membrane, Flow Cytometry

Journal: Scientific reports

Article Title: Orthogonal validation of PROTAC mediated degradation of the integral membrane proteins EGFR and c-MET.

doi: 10.1038/s41598-024-84217-2

Figure Lengend Snippet: Fig. 5. Degradation profile of ex19del EGFR in HCC827 cells. Cells were pre-treated as described in (Fig. 1) with EGF as RTK ligand and either PROTAC3 (A,C,E,G,I) or Gefitinib (B,D,F,H,J) was added for 24 h. Cells were then analysed by immunoblotting (A,B) with quantification (C,D), immunofluorescence for the whole cell (E,F) or plasma membrane only (G,H) or flow cytometry (I,J). Graphs show the mean SEM of at least 3 biological replicates. The dotted line indicates a value of 0.8. All details regarding statistical analyses and p-values are provided in the (Supplementary Table 4).

Article Snippet: For starvation experiments, cells were washed twice in PBS and grown in respective growth media without FBS for 6–8 h. Subsequently media were exchanged for full growth media and cells were treated with SJF8240 (Tocris, 7266), Foretinib (Stratech, ORB322222), PROTAC3 (Tocris, 7258) or Gefitinib (AstraZeneca).

Techniques: Western Blot, Immunofluorescence, Clinical Proteomics, Membrane, Flow Cytometry

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Accumulation of PNPLA3 on lipid droplets is the basis of associated hepatic steatosis

doi: 10.1073/pnas.1901974116

Figure Lengend Snippet: PROTAC-mediated degradation of Halo-PNPLA3(148M) reduces liver TGs. ( A ) A schematic of the experiment. Adapted with permission from ref. . Copyright 2015 American Chemical Society. ( B ) Recombinant AAVs were used to express Halo-tagged PNPLA3 constructs under the control of a liver-specific promoter (thyroxine-binding globulin). Female mice ( n = 3 to 7 per group, aged 12 wk) were injected with AAVs (1.25 × 10 11 GCs) expressing Halo alone, Halo-PNPLA3(WT), or Halo-PNPLA3(148M). After 2 wk on an HSD, the mice were treated with vehicle alone (2.5% DMSO in 0.9% NaCl) or PROTAC3 (4.8 mg/kg) for 2 wk (three doses per wk) and then killed after 3 d of dietary synchronization. Livers were harvested for lipid analysis and for isolation of LDs as described in SI Appendix, Methods . Bars represent mean ± SEM values. * P < 0.05, *** P < 0.001.

Article Snippet: Mice were fed an HSD for 2 wk and then treated with PROTAC3 (Promega; CS2072A01; 479 μg per vial) as described ( ).

Techniques: Recombinant, Construct, Control, Binding Assay, Injection, Expressing, Isolation