Figure 6 from TNG961 Is a Selective Oral HBS1L Molecular Glue Degrader for the Treatment of FOCAD-Deleted Cancers
Year: 2026
Type: other
Abstract: TNG961 induces translational arrest and activates ER stress and the UPR selectively in FOCAD-null cells. A, Translation assessed by puromycin incorporation in FOCAD isogenic MIAPACA2 cells after 48 hours of treatment with the indicated concentrations of TNG961. Puromycin immunoblot intensity was normalized to Ponceau staining to control for protein loading. Bars show mean ± SD (N = 3 biological replicates). **, P < 0.01 by two-way ANOVA with Sidak multiple comparisons test (vs. DMSO). Representative blots are shown in Supplementary Fig. S5A. B, HBS1L, IFRD1, and vinculin protein levels assessed by Simple Western in FOCAD isogenic MIAPACA2 cells after 48 hours of treatment with the indicated concentrations of TNG961. Representative images are from N = 2 biological replicates. C, ATF4 and vinculin protein levels assessed by Western blot in FOCAD isogenic MIAPACA2 cells after 48 hours of treatment with the indicated concentrations of TNG961. Tm, tunicamycin (positive control for ATF4 induction). Representative images are from N = 2 biological replicates. D, Spliced XBP1 (XBP1s) and actin protein levels assessed by Western blot in MIAPACA2 cells expressing a doxycycline (DOX)-inducible FOCAD cDNA (−DOX, FOCAD not expressed/FOCAD-null; +DOX, FOCAD expressed/FOCAD-reconstituted) after 72 hours of treatment with the indicated concentrations of TNG961. Representative images are from N = 2 biological replicates. E, qPCR analysis of CHOP, TRIB3, and GADD34 mRNA levels in FOCAD isogenic MIAPACA2 cells after 48 hours of treatment with the indicated concentrations of TNG961. Bars show mean ± SD (N = 4 biological replicates). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by one-way ANOVA with Dunnett multiple comparisons test (vs. DMSO). ANOVA P values (FOCAD null): CHOP P = 0.0044, TRIB3 P < 0.0001, GADD34 P = 0.0016; ANOVA P values (FOCAD reconstituted): CHOP P = 0.4290, TRIB3 P = 0.0320, GADD34 P = 0.0108.
Authors: Hilary E. Nicholson, Douglas A. Whittington, Frank J. Bruzzese, Katherine Lazarides, Lauren Catherine M. Martires, Matthew R. Tonini, Helena N. Jenkins, Minjie Zhang, Preksha Shahagadkar, Charlotte B. Pratt, Kimberly J. Briggs, Patrick McCarren, Alice W. Tsai, Madhavi Bandi, Chengyin Min, Alan Huang, Hongxiang Zhang, Samuel R. Meier, Binzhang Shen, Yi Yu, Colin Liang, Yong Liu, Teng Teng, John Zhang, Adam Crystal, William D. Mallender, Xinyuan Edward Wu, John P. Maxwell, Jannik N. Andersen
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W7204939193