Journal: bioRxiv

Article Title: Tau-induced mitochondrial reverse electron transport drives neurodegeneration

doi: 10.64898/2026.04.04.716514

Figure Lengend Snippet: a, Representative WB and quantification showing effect of VDAC, Hsp70, and Hsp90 inhibitor treatment on tau entry into mitochondria in the in vitro import assay. Mitochondrial p-tau level is normalized with the C-II protein SDHA. b , RET-ROS measurements after p-tau import into control or inhibitor-treated mitochondria. c , Representative WB and quantification showing total PHF-1 tau level in the in vitro import assay mixture. Total p-tau level is normalized with actin, which is known to be associated with mitochondria. d , RET-ROS measurements in control or inhibitor-treated mitochondria without p-tau import. e , WBs and quantification showing knockdown efficiency by VDAC1, Hsp70, and Hsp90 siRNAs. f , Representative WBs and quantification showing the effect of VDAC1, Hsp70, and Hsp90 siRNAs on tau entry into mitochondria in the in vitro import assay and total PHF-1 tau level in the in vitro import assay. g , h , RET-ROS measurements in mitochondria from control or siRNA treated cells with ( g ) or without ( h ) p-tau import into mitochondria. All data are means ± SEM; statistical significance was determined by one-way ANOVA with Dunnett’s multiple test ( a, b, c, d, f, g, h ), or two-tailed unpaired Student’s t test ( e ). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Each data point in a - h represents an independent experimental repeat.

Article Snippet: Newly emerged flies were treated with VDAC inhibitor DIDS (MedChemExpress #HY-D0086, 100 μM), Hsp70 inhibitor PES-CI (Sigma-Aldrich #5310670001, 60 μM), or Hsp90 inhibitor Geldanamycin (MedChemExpress #HY-15230, 10 μM) for 2-3 weeks.

Techniques: In Vitro, Control, Knockdown, Two Tailed Test

Journal: bioRxiv

Article Title: Tau-induced mitochondrial reverse electron transport drives neurodegeneration

doi: 10.64898/2026.04.04.716514

Figure Lengend Snippet: a, Representative WBs and quantification showing effect of VDAC, Hsp70, and Hsp90 inhibitors on the mitochondrial level of PHF-1 tau and total PHF-1 tau in the elav-GS>tau-R406W flies. In this GeneSwitch inducible model, tau-R406W expression is tightly controlled by the addition of RU486 to the fly food. Mitochondrial fractions or total cell lysates were used for WB, and mitochondrial or total PHF-1 p-tau was normalized by SDHA or actin. b , RET-ROS measurements in the mitochondria from control or inhibitor treated elav-GS>tau-R406W flies after tau induction by RU486. c - e , aversive taste memory assays in VDACi ( c ), Hsp70i ( d ), and Hsp90i ( e ) treated elav-GS>tau-R406W flies after tau induction by RU486. f , Climbing activity assay in VDACi, Hsp70i, and Hsp90i treated elav-GS>tau-R406W flies after tau induction by RU486. g , h , RET ( g ) and aversive taste memory ( h ) assays of control flies without tau transgene expression. All data are means ± SEM; statistical significance was determined by one-way ANOVA with Dunnett’s multiple test ( a, b, f, g ), or group analysis using multiple t test with Sidak-Bonferroni multiple comparison ( c , d , e , h ). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Each data point represents independent experimental repeat. Three sets of flies with 10-12 flies in each set were used for the behavioral assays.

Article Snippet: Newly emerged flies were treated with VDAC inhibitor DIDS (MedChemExpress #HY-D0086, 100 μM), Hsp70 inhibitor PES-CI (Sigma-Aldrich #5310670001, 60 μM), or Hsp90 inhibitor Geldanamycin (MedChemExpress #HY-15230, 10 μM) for 2-3 weeks.

Techniques: Expressing, Control, Activity Assay, Comparison

Journal: bioRxiv

Article Title: Tau-induced mitochondrial reverse electron transport drives neurodegeneration

doi: 10.64898/2026.04.04.716514

Figure Lengend Snippet: a , Effect of H 2 O 2 and FK866 on the viability of tau-P301L hiPSC neurons and isogenic wildtype controls after 24h treatment, and the rescuing effect of CPT. b , Effect of H 2 O 2 and FK866 co-treatment on the viability of control and tau KD hiPSC neurons and the rescuing effect of CPT. c, d , Effect of DES treatment on the viability of control and tau KD hiPSC neurons ( c ) and the rescuing effect of CPT in DES (20 mM) treated control hiPSC neurons ( d ). e - g , WB and quantification showing the effect of inhibiting VDAC, Hsp70, or Hsp90 on PHF-1 tau entry into mitochondria without affecting total PHF-1 tau levels ( e ), and quantification of the effect of inhibitor treatment on RET activity ( f ) and stress sensitivity ( g ) of APP hiPSC neurons. h , Measurement of RET ROS and NAD + /NADH in purified mitochondria from control and tau-WT-EGFP or tau-S2A-EGFP-transfected normal hiPSC neurons. i , Representative images and quantification of p-S262 tau in tau-WT-EGFP or tau-S2A-EGFP transfected control hiPSC neurons with or without DES treatment. j , Measurement of RET ROS and NAD + /NADH in purified mitochondria from EGFP or MKI-EGFP transduced APP hiPSC neurons. k , Representative images and quantification of p-S262 tau in EGFP or MKI-EGFP transfected APP hiPSC neurons. l, m , Immunoblots ( l ) and quantification ( m ) showing effects of the various treatments on normalized levels of p-tau species in APP hiPSC neurons. n, o , Immunoblots ( n ) and quantification ( o ) showing effect of DES or DES/CPT co-treatment on normalized levels of p-tau species in control hiPSC neurons. All data are means ± SEM; statistical significance was determined by one-way ANOVA with Tukey’s post hoc test ( a, b, d, g, h, i, o ), two-way ANOVA with Tukey’s post hoc test ( c ), two-tailed unpaired Student’s t test ( j, k ), or one-way ANOVA with Dunnett’s multiple test ( e, f , m ). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 ( a, b, c, d, g : n=6/group from 3 biological replicates and 2 wells/experiment; e, f, h, j, m, o : n=3 biological replicates; i, k : n=3 biological replicates, and each data point represents an average of 6 cells/experiment).

Article Snippet: Newly emerged flies were treated with VDAC inhibitor DIDS (MedChemExpress #HY-D0086, 100 μM), Hsp70 inhibitor PES-CI (Sigma-Aldrich #5310670001, 60 μM), or Hsp90 inhibitor Geldanamycin (MedChemExpress #HY-15230, 10 μM) for 2-3 weeks.

Techniques: Control, Activity Assay, Purification, Transfection, Western Blot, Two Tailed Test

Journal: bioRxiv

Article Title: Tau-induced mitochondrial reverse electron transport drives neurodegeneration

doi: 10.64898/2026.04.04.716514

Figure Lengend Snippet: a, Representative WBs and quantification showing effect of VDAC1, Hsp70, and Hsp90 lenti-shRNAs on the expression of the target proteins. b , Representative WBs and quantification showing effect of VDAC1, Hsp70, and Hsp90 lenti-shRNAs on the levels of mitochondrially localized PHF-1 tau and total PHF-1 tau in APP hiPSC neurons. Mitochondrial fractions or total cell lysates were used for WB, and mitochondrial or total PHF-1 p-tau was normalized by SDHA or actin. c , RET-ROS measurements in mitochondria from control or VDAC1, Hsp70, and Hsp90 lenti-shRNA treated APP hiPSC neurons. d , e , RET ( d ) and stress sensitivity ( e ) assays of control iPSC neurons treated with VDAC1, Hsp70, and Hsp90 inhibitors. All data are means ± SEM; statistical significance was determined by two-tailed unpaired Student’s t test ( a ) or one-way ANOVA with Dunnett’s multiple test ( b-e ). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001 ( e : n=6/group from 3 biological replicates and 2 wells/experiment).

Article Snippet: Newly emerged flies were treated with VDAC inhibitor DIDS (MedChemExpress #HY-D0086, 100 μM), Hsp70 inhibitor PES-CI (Sigma-Aldrich #5310670001, 60 μM), or Hsp90 inhibitor Geldanamycin (MedChemExpress #HY-15230, 10 μM) for 2-3 weeks.

Techniques: Expressing, Control, shRNA, Two Tailed Test

Journal: Cancers

Article Title: Targeting HSP90 with Ganetespib to Induce CDK1 Degradation and Promote Cell Death in Hepatoblastoma.

doi: 10.3390/cancers17081341

Figure Lengend Snippet: Figure 1. (A) A heat map showing the drug responses of 17 liver tumor models and 4 non-cancerous controls towards 5 HSP90 inhibitors (pimi)tespib, (gelda)namycin, (lumi)nespib, (tane)spimicin, and (gane)tespib as well as the standard-of-care drugs (carbo)platin, (cis)platin, and (dox)orubicin. Corresponding Z-scores and therapeutic window scores (TWS) are depicted as box-with-whiskers and lollipop plots, respectively. (B) Viability curves of liver tumor models towards 10 increasing concentrations of ganetespib (5 nm–100 µM). Dark pink and blue curves represent the mean response of the tumor models and healthy controls, respectively. (C) IC50 values of ganetespib-treated cell lines. (D) Principal component analysis of 83 compounds in our in vitro drug-testing platform. The drugs positioned on the right-hand side of the dashed line, which represents the PC1 of cisplatin, were considered ineffective. (E) Western blot analysis showing the HSP90 protein expression levels in two normal liver tissues and four hepatoblastoma models. Beta actin (ACTB) was used as a loading control. Bar graph shows the mean relative band density of two independent experiments ± SD. (F) Enrichment plot of gene ontology obtained from gene set enrichment analysis (GSEA) of 53 HB patients versus 14 non-cancerous livers from the publicly available GSE131329 data set. (G) Comparative RNA expression analysis of HSP90 in 53 HB patients and 14 non-cancerous liver samples from the GSE131329 data set, utilizing R2 Genomics Analysis and Visualization Platform. (H) Kaplan-Meier curves displaying overall survival (OS) and event-free survival (EFS) probabilities for patients with HB of the GSE131329 data set with either high (n = 27) or low (n = 25) HSP90 expression. Median expression was considered as cutout, and the log-rank Mantel–Cox test was used to calculate significance. **** p < 0.0001.

Article Snippet: MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide) assays were performed on 5 × 105 cells per well in a 96-well-plate after treating cells for 48 h with ten increasing doses ranging from 5 nM to 100 μM of carboplatin, cisplatin, and doxorubicin as well as the HSP90 inhibitors geldanamycin, luminespib, pimitespib, tanespimicin, and ganetespib (all from Selleck Chemicals GmbH, Houston, TX, USA) as described earlier [24].

Techniques: In Vitro, Western Blot, Expressing, Control, RNA Expression

Journal: Cancers

Article Title: Targeting HSP90 with Ganetespib to Induce CDK1 Degradation and Promote Cell Death in Hepatoblastoma.

doi: 10.3390/cancers17081341

Figure Lengend Snippet: Figure 4. (A) Western blot analysis (left) and the bar graphs with corresponding density calculation of the bands (right) showing alterations in heat shock protein 90 (HSP90) protein levels upon increasing concentrations of (gane)tespib for 24 h. Glycerinaldehyd-3-phosphat-dehydrogenase (GAPDH) was used as loading control. (B) Schematic illustration of recruitment of a client by HSP90. (C) KEGG pathway enrichment scores of 767 significantly upregulated (p < 0.05, fold > 2) genes from RNA sequencing analysis of GSE131329 data set. (D) Enrichment plot showing the KEGG category cell cycle, obtained from gene set enrichment analysis (GSEA) of the GSE131329 data set. (E) Comparative RNA expression analysis of HSP90 clients from the GSE131329 data set, utilizing R2 Genomics Analysis and Visualization Platform. Statistics were calculated using a two-tailed unpaired Student’s t-test, with *** p < 0.001, **** p < 0.0001. (F) Western blot showing the expression of cyclin-dependent kinase 1 (CDK1), polo-like kinase 1 (PLK1), and aurora kinase A (AURKA) in non-cancerous liver tissue and hepatoblastoma models. Beta actin (ACTB) and GAPDH served as loading controls. (G) Correlation of gene expression between CDK1, PLK1, and AURKA with HSP90 of the GSE131329 data. Two-tailed Pearson test was performed, and correlation coefficient was calculated.

Article Snippet: MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide) assays were performed on 5 × 105 cells per well in a 96-well-plate after treating cells for 48 h with ten increasing doses ranging from 5 nM to 100 μM of carboplatin, cisplatin, and doxorubicin as well as the HSP90 inhibitors geldanamycin, luminespib, pimitespib, tanespimicin, and ganetespib (all from Selleck Chemicals GmbH, Houston, TX, USA) as described earlier [24].

Techniques: Western Blot, Control, RNA Sequencing, RNA Expression, Two Tailed Test, Expressing, Gene Expression

Journal: The EMBO Journal

Article Title: Pharmacological CDK4/6 inhibition promotes vulnerability to lysosomotropic agents in breast cancer

doi: 10.1038/s44318-025-00371-x

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Geldanamycin (HSP90 inhibitor) , Cayman Chemical , 13355.

Techniques: Mouse Assay, Recombinant, Plasmid Preparation, Sequencing, CellTox Assay, Cytotoxicity Assay, Proliferation Assay, Staining, Transfection, Reverse Transcription, Software