rabbit anti pdi  (Cell Signaling Technology Inc)

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: Chronic cold exposure induces plasticity of mitochondrial calcium uptake in beige and brown fat of UCP1-deficient mice

doi: 10.64898/2026.03.16.712209

Figure Lengend Snippet: A. Swelling of BAT mitochondria from WT and Ucp1 -/- mice upon 3 weeks of cold exposure. B. Swelling of mitochondria from liver of WT mice kept at RT for comparison. C. Immunoblots of proteins implicated in ERMCs: GRP75, IP3R, VDAC, and cyclophilin D (CyD). D. Quantified protein expression. Calnexin, TOM20 and Na/K ATPase were used for normalization per ER biomass, mitochondrial biomass, and total extract, respectively. E. Representative image of PLA for IP3R–VDAC in BAT of WT and Ucp1 -/- mice exposed to 6°C for 3 weeks. F. Quantification of the contact sites from E . n = 4-6 mice. Data shown as mean ± sem. Unpaired t-test.

Article Snippet: Lysates were resolved by sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE), transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore), and probed with antibodies against Na + /K + -ATPase (Abcam, ab76020, GR3237646-13, 1/10,000), TOMM20 (Sigma Prestige, HPA011562, 1/500), MCU (Cell Signaling Technology, 14997, 1/100), UCP1 (Abcam, ab10983, 1/1000), NCLX (Proteintech, 21430-1-AP, 1/500), TMBIM5 (Proteintech, 16296-1-AP, 1/500), TMEM65 (Invitrogen, PA5-112762, 1/500), GRP75 (Abcam, ab53098, 1/500), Calnexin (BD Transduction Laboratories, 610523, 1/1000), VDAC (Cell Signaling Technology, 4661, 1/2000), CyD (Proteintech, 18466-1-AP, 1/1000), OXPHOS (Abcam, ab110413, 1/500), Vinculin (1:10000; V9131; Sigma-Aldrich), NDUFB6 (1:5000; #6510832 Thermo Fisher Scientific), mtCOI (1:10000; #10444035; Thermo Fisher Scientific), ATP5A (1:1,000; #439800; Thermo Fisher Scientific), SDHA (1:1,000, #459200, Thermo Fisher Scientific), SDHB (1:1,000, #ab14714, Abcam), UQCRC2 (1:1,000, #14742-1-AP; Proteintech), ATPIF1 (1:1,000, #13268S; Cell Signaling Technology), TOMM20 Alexa Fluor 488 (1:1,000; #ab205486; Abcam), and cytochrome c (1:5,000, #ab110325, Abcam).

Techniques: Comparison, Western Blot, Expressing