anti hsp90 Search Results


93
StressMarq anti hsp90
Anti Hsp90, supplied by StressMarq, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Rockland Immunochemicals anti acetylated hsp90
Western blot analysis of <t>acetyl-HSP90,</t> HSP90, and PAH protein expression in liver tissues from sham, treated, and wild-type groups following 12 weeks of TSA treatment. ( A ) Expression of acetyl-HSP90, HSP90, PAH, and GAPDH (used as a loading control). ( B ) Quantitative analysis of acetyl-HSP90 relative to HSP90 protein levels. ( C ) Quantitative analysis of PAH relative to GAPDH protein levels. The expression of PAE was quantified using the Image J software (Version 1.49) and expressed as arbitrary units (a.u.). Results are presented as mean ± SEM, with statistical significance indicated. ****P<0.0001.
Anti Acetylated Hsp90, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/HSP90+K294+Antibody/pmc12748783-156-3-6
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Rockland Immunochemicals anti aha1
Western blot analysis of <t>acetyl-HSP90,</t> HSP90, and PAH protein expression in liver tissues from sham, treated, and wild-type groups following 12 weeks of TSA treatment. ( A ) Expression of acetyl-HSP90, HSP90, PAH, and GAPDH (used as a loading control). ( B ) Quantitative analysis of acetyl-HSP90 relative to HSP90 protein levels. ( C ) Quantitative analysis of PAH relative to GAPDH protein levels. The expression of PAE was quantified using the Image J software (Version 1.49) and expressed as arbitrary units (a.u.). Results are presented as mean ± SEM, with statistical significance indicated. ****P<0.0001.
Anti Aha1, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/AHA1+Antibody/pmc03689215-98-0-4
Average 90 stars, based on 1 article reviews
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91
StressMarq hsp90
Identification of the heat shock proteins HSP70 and <t>HSP90</t> as putative ChAT protein-interactors by proximity-dependent biotin identification (BioID). (A) Optimization of BioID in HEK293 cells expressing wild-type or P17A/P19A-ChAT fused to the HA-tagged promiscuous biotin ligase BirA-R118G (BirA * ). Control cells were transfected with empty vector or plasmids encoding either untagged ChAT or BirA * . Cells were treated for 24 h with either 50 μM biotin to facilitate proximity-dependent biotinylation of ChAT-interacting cellular proteins or with vehicle-control (water). Biotinylated proteins were isolated from cell lysates by streptavidin pull-downs (PD: Strep) and immunoblotted as indicated ( n = 2). (B) Identification of HSP70 and HSP90 as ChAT proximally-interacting proteins. Streptavidin PD samples prepared from biotin-treated HEK293 cells expressing wild-type-ChAT-BirA * or P17A/P19A-ChAT-BirA * fusion proteins were resolved and visualized on a silver-stained SDS-PAGE gels. Two proteins (~70 and ~90 kDa) that were enriched in samples expressing P17A/P19A-ChAT-BirA * were identified by MALDI-TOF-MS or LC-ESI-MS/MS as HSP70 and HSP90, respectively. Control cells were transfected to express untagged wild-type ChAT ( n = 1). (C) Confirmation of endogenous HSP70 and HSP90 as putative ChAT-interacting proteins by immunoblotting of streptavidin PD samples prepared from biotin-treated HEK293 cells expressing HA-tagged wild-type-ChAT-BirA * or P17A/P19A-ChAT-BirA * . Control cells were transfected with empty vector or vector encoding untagged ChAT ( n = 4).
Hsp90, supplied by StressMarq, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/Anti-HSP90+Antibody/pmc05733026-98-53-55
Average 91 stars, based on 1 article reviews
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StressMarq cat smc
Identification of the heat shock proteins HSP70 and <t>HSP90</t> as putative ChAT protein-interactors by proximity-dependent biotin identification (BioID). (A) Optimization of BioID in HEK293 cells expressing wild-type or P17A/P19A-ChAT fused to the HA-tagged promiscuous biotin ligase BirA-R118G (BirA * ). Control cells were transfected with empty vector or plasmids encoding either untagged ChAT or BirA * . Cells were treated for 24 h with either 50 μM biotin to facilitate proximity-dependent biotinylation of ChAT-interacting cellular proteins or with vehicle-control (water). Biotinylated proteins were isolated from cell lysates by streptavidin pull-downs (PD: Strep) and immunoblotted as indicated ( n = 2). (B) Identification of HSP70 and HSP90 as ChAT proximally-interacting proteins. Streptavidin PD samples prepared from biotin-treated HEK293 cells expressing wild-type-ChAT-BirA * or P17A/P19A-ChAT-BirA * fusion proteins were resolved and visualized on a silver-stained SDS-PAGE gels. Two proteins (~70 and ~90 kDa) that were enriched in samples expressing P17A/P19A-ChAT-BirA * were identified by MALDI-TOF-MS or LC-ESI-MS/MS as HSP70 and HSP90, respectively. Control cells were transfected to express untagged wild-type ChAT ( n = 1). (C) Confirmation of endogenous HSP70 and HSP90 as putative ChAT-interacting proteins by immunoblotting of streptavidin PD samples prepared from biotin-treated HEK293 cells expressing HA-tagged wild-type-ChAT-BirA * or P17A/P19A-ChAT-BirA * . Control cells were transfected with empty vector or vector encoding untagged ChAT ( n = 4).
Cat Smc, supplied by StressMarq, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/Anti-HSP90+alpha+Antibody/pmc06948982-485-39-43
Average 92 stars, based on 1 article reviews
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92
StressMarq mouse anti hsp90
No significant change in expression of HSP70 isoforms, HSP40, <t>HSP90</t> or HSPB1 in FUS R521G cortex or with drug treatments. FUS R521G mice were treated daily i.p . for 28 days beginning on P60 with vehicle, 20 ​mg/kg RGFP963, 10 ​mg/kg RGFP963), 20 ​mg/kg arimoclomol or the combination. Western blots of homogenates of cortex were probed with antibodies smc-100 (HSPA1A), SPA812 (inducible and constitutive HSP70), smc-151 (HSPA8), ADI-SPA-400 (HSP40), smc-149 (HSP90) or sc-1049 (HSPB1) and normalized to GAPDH or actin. Top row: Levels of HSPs were unchanged in cortex of FUS R521G compared to control mice. Rows 2–5: Levels were not affected by any drug treatments. Data are shown as mean ​± ​SD (3–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.
Mouse Anti Hsp90, supplied by StressMarq, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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StressMarq mouse anti p23 monoclonal antibody
No significant change in expression of HSP70 isoforms, HSP40, <t>HSP90</t> or HSPB1 in FUS R521G cortex or with drug treatments. FUS R521G mice were treated daily i.p . for 28 days beginning on P60 with vehicle, 20 ​mg/kg RGFP963, 10 ​mg/kg RGFP963), 20 ​mg/kg arimoclomol or the combination. Western blots of homogenates of cortex were probed with antibodies smc-100 (HSPA1A), SPA812 (inducible and constitutive HSP70), smc-151 (HSPA8), ADI-SPA-400 (HSP40), smc-149 (HSP90) or sc-1049 (HSPB1) and normalized to GAPDH or actin. Top row: Levels of HSPs were unchanged in cortex of FUS R521G compared to control mice. Rows 2–5: Levels were not affected by any drug treatments. Data are shown as mean ​± ​SD (3–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.
Mouse Anti P23 Monoclonal Antibody, supplied by StressMarq, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/Anti-p23+Antibody/pmc03477583-71-13-19
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93
StressMarq hsp90 antibody
No significant change in expression of HSP70 isoforms, HSP40, <t>HSP90</t> or HSPB1 in FUS R521G cortex or with drug treatments. FUS R521G mice were treated daily i.p . for 28 days beginning on P60 with vehicle, 20 ​mg/kg RGFP963, 10 ​mg/kg RGFP963), 20 ​mg/kg arimoclomol or the combination. Western blots of homogenates of cortex were probed with antibodies smc-100 (HSPA1A), SPA812 (inducible and constitutive HSP70), smc-151 (HSPA8), ADI-SPA-400 (HSP40), smc-149 (HSP90) or sc-1049 (HSPB1) and normalized to GAPDH or actin. Top row: Levels of HSPs were unchanged in cortex of FUS R521G compared to control mice. Rows 2–5: Levels were not affected by any drug treatments. Data are shown as mean ​± ​SD (3–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.
Hsp90 Antibody, supplied by StressMarq, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/Anti-HSP90+Antibody/10__1523_slash_jneurosci__5634___11__2012-106-25-27
Average 93 stars, based on 1 article reviews
hsp90 antibody - by Bioz Stars, 2026-11
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92
StressMarq rabbit antihsp90
No significant change in expression of HSP70 isoforms, HSP40, <t>HSP90</t> or HSPB1 in FUS R521G cortex or with drug treatments. FUS R521G mice were treated daily i.p . for 28 days beginning on P60 with vehicle, 20 ​mg/kg RGFP963, 10 ​mg/kg RGFP963), 20 ​mg/kg arimoclomol or the combination. Western blots of homogenates of cortex were probed with antibodies smc-100 (HSPA1A), SPA812 (inducible and constitutive HSP70), smc-151 (HSPA8), ADI-SPA-400 (HSP40), smc-149 (HSP90) or sc-1049 (HSPB1) and normalized to GAPDH or actin. Top row: Levels of HSPs were unchanged in cortex of FUS R521G compared to control mice. Rows 2–5: Levels were not affected by any drug treatments. Data are shown as mean ​± ​SD (3–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.
Rabbit Antihsp90, supplied by StressMarq, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/Anti-HSP90+Antibody/pm22734125-61-33-35
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StressMarq mouse anti human hsp 90 alpha antibody
No significant change in expression of HSP70 isoforms, HSP40, <t>HSP90</t> or HSPB1 in FUS R521G cortex or with drug treatments. FUS R521G mice were treated daily i.p . for 28 days beginning on P60 with vehicle, 20 ​mg/kg RGFP963, 10 ​mg/kg RGFP963), 20 ​mg/kg arimoclomol or the combination. Western blots of homogenates of cortex were probed with antibodies smc-100 (HSPA1A), SPA812 (inducible and constitutive HSP70), smc-151 (HSPA8), ADI-SPA-400 (HSP40), smc-149 (HSP90) or sc-1049 (HSPB1) and normalized to GAPDH or actin. Top row: Levels of HSPs were unchanged in cortex of FUS R521G compared to control mice. Rows 2–5: Levels were not affected by any drug treatments. Data are shown as mean ​± ​SD (3–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.
Mouse Anti Human Hsp 90 Alpha Antibody, supplied by StressMarq, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/Anti-HSP90+alpha+Antibody/pmc11323689-108-0-36
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Boster Bio ahsa1
Quantitative expression profiles of UPP1, LACTB, and <t>AHSA1</t> proteins in omics analysis. (A–C) Protein quantification values of UPP1, LACTB, and AHSA1 in PC vs para-PC group; (D–F) Protein quantification values of UPP1, LACTB, and AHSA1 in PD-PC vs MD-PC group. * P < 0.05. PD: Poorly differentiated; PC: Pancreatic cancer; MD: Moderately differentiated; UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1.
Ahsa1, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/Anti-AHA1+Monoclonal+Antibody/pmc12452116-187-15-17
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StressMarq hsp82 antibody
Quantitative expression profiles of UPP1, LACTB, and <t>AHSA1</t> proteins in omics analysis. (A–C) Protein quantification values of UPP1, LACTB, and AHSA1 in PC vs para-PC group; (D–F) Protein quantification values of UPP1, LACTB, and AHSA1 in PD-PC vs MD-PC group. * P < 0.05. PD: Poorly differentiated; PC: Pancreatic cancer; MD: Moderately differentiated; UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1.
Hsp82 Antibody, supplied by StressMarq, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+hsp90/Anti-HSP90+alpha%2Fbeta+Antibody/pmc06561935__41467_2019_10463_MOESM1_ESM-47-20-22
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Image Search Results


Western blot analysis of acetyl-HSP90, HSP90, and PAH protein expression in liver tissues from sham, treated, and wild-type groups following 12 weeks of TSA treatment. ( A ) Expression of acetyl-HSP90, HSP90, PAH, and GAPDH (used as a loading control). ( B ) Quantitative analysis of acetyl-HSP90 relative to HSP90 protein levels. ( C ) Quantitative analysis of PAH relative to GAPDH protein levels. The expression of PAE was quantified using the Image J software (Version 1.49) and expressed as arbitrary units (a.u.). Results are presented as mean ± SEM, with statistical significance indicated. ****P<0.0001.

Journal: Scientific Reports

Article Title: Therapeutic effect of histone deacetylase 6 inhibitor for a mouse model of phenylketonuria

doi: 10.1038/s41598-025-29143-7

Figure Lengend Snippet: Western blot analysis of acetyl-HSP90, HSP90, and PAH protein expression in liver tissues from sham, treated, and wild-type groups following 12 weeks of TSA treatment. ( A ) Expression of acetyl-HSP90, HSP90, PAH, and GAPDH (used as a loading control). ( B ) Quantitative analysis of acetyl-HSP90 relative to HSP90 protein levels. ( C ) Quantitative analysis of PAH relative to GAPDH protein levels. The expression of PAE was quantified using the Image J software (Version 1.49) and expressed as arbitrary units (a.u.). Results are presented as mean ± SEM, with statistical significance indicated. ****P<0.0001.

Article Snippet: The antibodies used anti-acetylated HSP90 (600-401-981, Rockland Immunochemical, 1:500), anti-HSP90 (#4877, Cell Signaling, 1:1,000), and PAH (sc-271258, Santa Cruz, 1:2,000).

Techniques: Western Blot, Expressing, Control, Software

Mechanism of TSA in enhancing HSP90 acetylation and stabilizing misfolded PAH protein to improve phenylalanine metabolism.

Journal: Scientific Reports

Article Title: Therapeutic effect of histone deacetylase 6 inhibitor for a mouse model of phenylketonuria

doi: 10.1038/s41598-025-29143-7

Figure Lengend Snippet: Mechanism of TSA in enhancing HSP90 acetylation and stabilizing misfolded PAH protein to improve phenylalanine metabolism.

Article Snippet: The antibodies used anti-acetylated HSP90 (600-401-981, Rockland Immunochemical, 1:500), anti-HSP90 (#4877, Cell Signaling, 1:1,000), and PAH (sc-271258, Santa Cruz, 1:2,000).

Techniques:

Identification of the heat shock proteins HSP70 and HSP90 as putative ChAT protein-interactors by proximity-dependent biotin identification (BioID). (A) Optimization of BioID in HEK293 cells expressing wild-type or P17A/P19A-ChAT fused to the HA-tagged promiscuous biotin ligase BirA-R118G (BirA * ). Control cells were transfected with empty vector or plasmids encoding either untagged ChAT or BirA * . Cells were treated for 24 h with either 50 μM biotin to facilitate proximity-dependent biotinylation of ChAT-interacting cellular proteins or with vehicle-control (water). Biotinylated proteins were isolated from cell lysates by streptavidin pull-downs (PD: Strep) and immunoblotted as indicated ( n = 2). (B) Identification of HSP70 and HSP90 as ChAT proximally-interacting proteins. Streptavidin PD samples prepared from biotin-treated HEK293 cells expressing wild-type-ChAT-BirA * or P17A/P19A-ChAT-BirA * fusion proteins were resolved and visualized on a silver-stained SDS-PAGE gels. Two proteins (~70 and ~90 kDa) that were enriched in samples expressing P17A/P19A-ChAT-BirA * were identified by MALDI-TOF-MS or LC-ESI-MS/MS as HSP70 and HSP90, respectively. Control cells were transfected to express untagged wild-type ChAT ( n = 1). (C) Confirmation of endogenous HSP70 and HSP90 as putative ChAT-interacting proteins by immunoblotting of streptavidin PD samples prepared from biotin-treated HEK293 cells expressing HA-tagged wild-type-ChAT-BirA * or P17A/P19A-ChAT-BirA * . Control cells were transfected with empty vector or vector encoding untagged ChAT ( n = 4).

Journal: Frontiers in Molecular Neuroscience

Article Title: Chaperone-Mediated Regulation of Choline Acetyltransferase Protein Stability and Activity by HSC/HSP70, HSP90, and p97/VCP

doi: 10.3389/fnmol.2017.00415

Figure Lengend Snippet: Identification of the heat shock proteins HSP70 and HSP90 as putative ChAT protein-interactors by proximity-dependent biotin identification (BioID). (A) Optimization of BioID in HEK293 cells expressing wild-type or P17A/P19A-ChAT fused to the HA-tagged promiscuous biotin ligase BirA-R118G (BirA * ). Control cells were transfected with empty vector or plasmids encoding either untagged ChAT or BirA * . Cells were treated for 24 h with either 50 μM biotin to facilitate proximity-dependent biotinylation of ChAT-interacting cellular proteins or with vehicle-control (water). Biotinylated proteins were isolated from cell lysates by streptavidin pull-downs (PD: Strep) and immunoblotted as indicated ( n = 2). (B) Identification of HSP70 and HSP90 as ChAT proximally-interacting proteins. Streptavidin PD samples prepared from biotin-treated HEK293 cells expressing wild-type-ChAT-BirA * or P17A/P19A-ChAT-BirA * fusion proteins were resolved and visualized on a silver-stained SDS-PAGE gels. Two proteins (~70 and ~90 kDa) that were enriched in samples expressing P17A/P19A-ChAT-BirA * were identified by MALDI-TOF-MS or LC-ESI-MS/MS as HSP70 and HSP90, respectively. Control cells were transfected to express untagged wild-type ChAT ( n = 1). (C) Confirmation of endogenous HSP70 and HSP90 as putative ChAT-interacting proteins by immunoblotting of streptavidin PD samples prepared from biotin-treated HEK293 cells expressing HA-tagged wild-type-ChAT-BirA * or P17A/P19A-ChAT-BirA * . Control cells were transfected with empty vector or vector encoding untagged ChAT ( n = 4).

Article Snippet: Cells were washed with HBSS, formalin-fixed (4% paraformaldehyde in HBSS) for 15 min, permeabilized with 0.1% Triton X-100, blocked for 1 h in HBSS supplemented with 3% donkey serum, then finally incubated for 1 h with primary antibodies targeting ChAT (1:100; Chemicon, goat primary) together with either endogenous HSC70 (1:100; StressMarq, mouse primary), HSP90 (1:200; StressMarq, mouse primary) or CHIP (1:200; Santa Cruz, rabbit primary); all steps were performed at room temperature.

Techniques: Expressing, Transfection, Plasmid Preparation, Isolation, Staining, SDS Page, Tandem Mass Spectroscopy, Western Blot

Co-immunoprecipitation (co-IP) of ChAT with heat shock proteins HSC70, HSP70, and HSP90 is altered by mutation of N-terminal proline-rich motif in ChAT. (A) Immunoblots showing co-IP of ChAT with endogenous HSC70, HSP70 and HSP90 from HEK293 cells expressing either wild-type or P17A/P19A-ChAT. Control cells were transfected with empty vector. Using HEK293 cells, co-IP of P17A/P19A-ChAT with HSP70 (B) , HSP90 (C) and HSC70 (D) , respectively, is greater than that of wild-type ChAT ( *** p ≤ 0.001, Student's t -test, mean ± SEM, n = 4). (E) Co-IP of ChAT with endogenous HSC70 and HSP90 from mouse cholinergic SN56 cells expressing either wild-type or P17A/P19A-ChAT or CMS-related mutant proteins V18M- or A513T-ChAT. Control cells were transfected with empty vector. (F) Using SN56 cells, Co-IP of P17A/P19A-ChAT ( *** p ≤ 0.001) and V18M-ChAT ( * p ≤ 0.05), but not A531T-ChAT, with HSC70 is greater than that of wild-type ChAT (mean ± SEM, n = 5). (G) While there was a trend toward increased HSP90 interaction with P17A/P19A-ChAT ( p = 0.09), no significant differences were observed for HSP90 interaction with mutant ChAT compared to wild-type ChAT in SN56 cells (mean ± SEM, n = 5). Statistical analysis for (F) and (G) was performed by one-way ANOVA with Dunnett's post-hoc test. (H) Detection of in situ interactions of wild-type ChAT with endogenous HSC70 and HSP90 by proximity ligation assay (PLA) in SN56 cells. Formalin-fixed cells were first co-labeled with goat anti-ChAT together with either mouse anti-HSC70 or mouse anti-HSP90 primary antibodies, then incubated with oligonucleotide-linked secondary antibodies. Following DNA ligation and DNA amplification using the Duolink in Situ Orange Kit (Sigma), in situ ChAT-HSP interactions were imaged by confocal microscopy. Positive in situ ChAT-HSP interactions where visualized as fluorescent red dots while nuclei were stained with DAPI (blue). Control cells were either transfected with empty vector or had primary antibodies omitted from the assay (No 1° antibodies). Images are representative of 3 independent experiments; scale bars are 10 μm.

Journal: Frontiers in Molecular Neuroscience

Article Title: Chaperone-Mediated Regulation of Choline Acetyltransferase Protein Stability and Activity by HSC/HSP70, HSP90, and p97/VCP

doi: 10.3389/fnmol.2017.00415

Figure Lengend Snippet: Co-immunoprecipitation (co-IP) of ChAT with heat shock proteins HSC70, HSP70, and HSP90 is altered by mutation of N-terminal proline-rich motif in ChAT. (A) Immunoblots showing co-IP of ChAT with endogenous HSC70, HSP70 and HSP90 from HEK293 cells expressing either wild-type or P17A/P19A-ChAT. Control cells were transfected with empty vector. Using HEK293 cells, co-IP of P17A/P19A-ChAT with HSP70 (B) , HSP90 (C) and HSC70 (D) , respectively, is greater than that of wild-type ChAT ( *** p ≤ 0.001, Student's t -test, mean ± SEM, n = 4). (E) Co-IP of ChAT with endogenous HSC70 and HSP90 from mouse cholinergic SN56 cells expressing either wild-type or P17A/P19A-ChAT or CMS-related mutant proteins V18M- or A513T-ChAT. Control cells were transfected with empty vector. (F) Using SN56 cells, Co-IP of P17A/P19A-ChAT ( *** p ≤ 0.001) and V18M-ChAT ( * p ≤ 0.05), but not A531T-ChAT, with HSC70 is greater than that of wild-type ChAT (mean ± SEM, n = 5). (G) While there was a trend toward increased HSP90 interaction with P17A/P19A-ChAT ( p = 0.09), no significant differences were observed for HSP90 interaction with mutant ChAT compared to wild-type ChAT in SN56 cells (mean ± SEM, n = 5). Statistical analysis for (F) and (G) was performed by one-way ANOVA with Dunnett's post-hoc test. (H) Detection of in situ interactions of wild-type ChAT with endogenous HSC70 and HSP90 by proximity ligation assay (PLA) in SN56 cells. Formalin-fixed cells were first co-labeled with goat anti-ChAT together with either mouse anti-HSC70 or mouse anti-HSP90 primary antibodies, then incubated with oligonucleotide-linked secondary antibodies. Following DNA ligation and DNA amplification using the Duolink in Situ Orange Kit (Sigma), in situ ChAT-HSP interactions were imaged by confocal microscopy. Positive in situ ChAT-HSP interactions where visualized as fluorescent red dots while nuclei were stained with DAPI (blue). Control cells were either transfected with empty vector or had primary antibodies omitted from the assay (No 1° antibodies). Images are representative of 3 independent experiments; scale bars are 10 μm.

Article Snippet: Cells were washed with HBSS, formalin-fixed (4% paraformaldehyde in HBSS) for 15 min, permeabilized with 0.1% Triton X-100, blocked for 1 h in HBSS supplemented with 3% donkey serum, then finally incubated for 1 h with primary antibodies targeting ChAT (1:100; Chemicon, goat primary) together with either endogenous HSC70 (1:100; StressMarq, mouse primary), HSP90 (1:200; StressMarq, mouse primary) or CHIP (1:200; Santa Cruz, rabbit primary); all steps were performed at room temperature.

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Mutagenesis, Western Blot, Expressing, Transfection, Plasmid Preparation, In Situ, Proximity Ligation Assay, Labeling, Incubation, DNA Ligation, Amplification, Confocal Microscopy, Staining

Inhibition of HSP90 ATPase activity specifically reduces steady-state levels of mutant ChAT protein. Immunoblots from SN56 cells expressing wild-type (A) , P17A/P19A- (B) , V18M- (C) , or A513T-ChAT (D) that were treated for 24 h with 0.5–2 μM with 17-AAG, an inhibitor of HSP90 activity, or with DMSO-vehicle. (E) Treatment of cells with 17-AAG at concentrations up to 2 μM has no effect on the steady-state levels of wild-type ChAT protein. Conversely, steady-state protein levels of P17A/P19A- (F) , V18M- (G) , and A513T-ChAT (H) are reduced following treatment of cells with 17-AAG compared to vehicle-control ( *** p ≤ 0.001; one-way ANOVA with Dunnett's post-hoc test, mean ± SEM, n = 4). (I) Proteasome inhibition by co-treatment with 5 μM MG132 for 18 h attenuates the effects of inhibition of HSP90 (1 μM 17-AAG, 24 h) on ChAT steady-state protein levels in ChAT-expressing SN56 cells. Control cells were treated with DMSO-vehicle. Proteasome inhibition was validated by immunoblotting for the accumulation of ubiquitinated cellular proteins ( n = 4). (J) Lysosomal inhibition by co-treatment with 50 μM chloroquine (CQ) for 18 h did not prevent the effects of 17-AAG treatment (1 μM, 24 h) on steady-state ChAT protein levels in SN56 cells. Lysosomal inhibition was validated by immunoblotting for the accumulation of the lysosome-associated protein LC3B-II ( n = 3).

Journal: Frontiers in Molecular Neuroscience

Article Title: Chaperone-Mediated Regulation of Choline Acetyltransferase Protein Stability and Activity by HSC/HSP70, HSP90, and p97/VCP

doi: 10.3389/fnmol.2017.00415

Figure Lengend Snippet: Inhibition of HSP90 ATPase activity specifically reduces steady-state levels of mutant ChAT protein. Immunoblots from SN56 cells expressing wild-type (A) , P17A/P19A- (B) , V18M- (C) , or A513T-ChAT (D) that were treated for 24 h with 0.5–2 μM with 17-AAG, an inhibitor of HSP90 activity, or with DMSO-vehicle. (E) Treatment of cells with 17-AAG at concentrations up to 2 μM has no effect on the steady-state levels of wild-type ChAT protein. Conversely, steady-state protein levels of P17A/P19A- (F) , V18M- (G) , and A513T-ChAT (H) are reduced following treatment of cells with 17-AAG compared to vehicle-control ( *** p ≤ 0.001; one-way ANOVA with Dunnett's post-hoc test, mean ± SEM, n = 4). (I) Proteasome inhibition by co-treatment with 5 μM MG132 for 18 h attenuates the effects of inhibition of HSP90 (1 μM 17-AAG, 24 h) on ChAT steady-state protein levels in ChAT-expressing SN56 cells. Control cells were treated with DMSO-vehicle. Proteasome inhibition was validated by immunoblotting for the accumulation of ubiquitinated cellular proteins ( n = 4). (J) Lysosomal inhibition by co-treatment with 50 μM chloroquine (CQ) for 18 h did not prevent the effects of 17-AAG treatment (1 μM, 24 h) on steady-state ChAT protein levels in SN56 cells. Lysosomal inhibition was validated by immunoblotting for the accumulation of the lysosome-associated protein LC3B-II ( n = 3).

Article Snippet: Cells were washed with HBSS, formalin-fixed (4% paraformaldehyde in HBSS) for 15 min, permeabilized with 0.1% Triton X-100, blocked for 1 h in HBSS supplemented with 3% donkey serum, then finally incubated for 1 h with primary antibodies targeting ChAT (1:100; Chemicon, goat primary) together with either endogenous HSC70 (1:100; StressMarq, mouse primary), HSP90 (1:200; StressMarq, mouse primary) or CHIP (1:200; Santa Cruz, rabbit primary); all steps were performed at room temperature.

Techniques: Inhibition, Activity Assay, Mutagenesis, Western Blot, Expressing

Inhibition of HSC/HSP70 and HSP90 activity enhances ChAT ubiquitination and reduces cellular ChAT enzymatic activity. (A) Immunoblots following anti-ChAT IP from ChAT-expressing SN56 cells that were co-treated for 24 h with 40 μM VER-155008 and for the final 6 h with 20 μM MG132 to inhibit degradation of ubiquitinated ChAT. Control cells were transfected with empty vector and/or treated with DMSO-vehicle. Inhibition of HSC/HSP70 by VER-155008 treatment enhanced ChAT ubiquitination, where levels of ubiquitinated mutant ChAT, particularly P17A/P19A-ChAT, are greater than that of wild-type ChAT ( n = 3). (B) Immunoblots following anti-ChAT IP from ChAT-expressing SN56 cells that were co-treated for either 8 or 24 h with 1 μM 17-AAG and for the final 6 h with 20 μM MG132. Inhibition of HSP90 by treatment with 1 μM 17-AAG for 8 h, but not for 24 h, enhanced ChAT ubiquitination where levels of ubiquitinated mutant ChAT, particularly P17A/P19A-ChAT, are greater than that of wild-type ChAT ( n = 3). (C) Cellular activity of wild-type ChAT is reduced following treatment of SN56 cells for 24 h with either 40 μM VER-155008 or 1 μM 17-AAG compared to vehicle-treated (DMSO) cells ( ** p ≤ 0.01). Treatment with VER-155008 trended toward a greater reduction in ChAT activity compared to 17-AAG ( p = 0.074; one-way ANOVA with Tukey's post-hoc test, mean ± SEM, n = 3).

Journal: Frontiers in Molecular Neuroscience

Article Title: Chaperone-Mediated Regulation of Choline Acetyltransferase Protein Stability and Activity by HSC/HSP70, HSP90, and p97/VCP

doi: 10.3389/fnmol.2017.00415

Figure Lengend Snippet: Inhibition of HSC/HSP70 and HSP90 activity enhances ChAT ubiquitination and reduces cellular ChAT enzymatic activity. (A) Immunoblots following anti-ChAT IP from ChAT-expressing SN56 cells that were co-treated for 24 h with 40 μM VER-155008 and for the final 6 h with 20 μM MG132 to inhibit degradation of ubiquitinated ChAT. Control cells were transfected with empty vector and/or treated with DMSO-vehicle. Inhibition of HSC/HSP70 by VER-155008 treatment enhanced ChAT ubiquitination, where levels of ubiquitinated mutant ChAT, particularly P17A/P19A-ChAT, are greater than that of wild-type ChAT ( n = 3). (B) Immunoblots following anti-ChAT IP from ChAT-expressing SN56 cells that were co-treated for either 8 or 24 h with 1 μM 17-AAG and for the final 6 h with 20 μM MG132. Inhibition of HSP90 by treatment with 1 μM 17-AAG for 8 h, but not for 24 h, enhanced ChAT ubiquitination where levels of ubiquitinated mutant ChAT, particularly P17A/P19A-ChAT, are greater than that of wild-type ChAT ( n = 3). (C) Cellular activity of wild-type ChAT is reduced following treatment of SN56 cells for 24 h with either 40 μM VER-155008 or 1 μM 17-AAG compared to vehicle-treated (DMSO) cells ( ** p ≤ 0.01). Treatment with VER-155008 trended toward a greater reduction in ChAT activity compared to 17-AAG ( p = 0.074; one-way ANOVA with Tukey's post-hoc test, mean ± SEM, n = 3).

Article Snippet: Cells were washed with HBSS, formalin-fixed (4% paraformaldehyde in HBSS) for 15 min, permeabilized with 0.1% Triton X-100, blocked for 1 h in HBSS supplemented with 3% donkey serum, then finally incubated for 1 h with primary antibodies targeting ChAT (1:100; Chemicon, goat primary) together with either endogenous HSC70 (1:100; StressMarq, mouse primary), HSP90 (1:200; StressMarq, mouse primary) or CHIP (1:200; Santa Cruz, rabbit primary); all steps were performed at room temperature.

Techniques: Inhibition, Activity Assay, Western Blot, Expressing, Transfection, Plasmid Preparation, Mutagenesis

No significant change in expression of HSP70 isoforms, HSP40, HSP90 or HSPB1 in FUS R521G cortex or with drug treatments. FUS R521G mice were treated daily i.p . for 28 days beginning on P60 with vehicle, 20 ​mg/kg RGFP963, 10 ​mg/kg RGFP963), 20 ​mg/kg arimoclomol or the combination. Western blots of homogenates of cortex were probed with antibodies smc-100 (HSPA1A), SPA812 (inducible and constitutive HSP70), smc-151 (HSPA8), ADI-SPA-400 (HSP40), smc-149 (HSP90) or sc-1049 (HSPB1) and normalized to GAPDH or actin. Top row: Levels of HSPs were unchanged in cortex of FUS R521G compared to control mice. Rows 2–5: Levels were not affected by any drug treatments. Data are shown as mean ​± ​SD (3–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.

Journal: Neurotherapeutics

Article Title: Reversal of cognitive deficits in FUS R521G amyotrophic lateral sclerosis mice by arimoclomol and a class I histone deacetylase inhibitor independent of heat shock protein induction

doi: 10.1016/j.neurot.2024.e00388

Figure Lengend Snippet: No significant change in expression of HSP70 isoforms, HSP40, HSP90 or HSPB1 in FUS R521G cortex or with drug treatments. FUS R521G mice were treated daily i.p . for 28 days beginning on P60 with vehicle, 20 ​mg/kg RGFP963, 10 ​mg/kg RGFP963), 20 ​mg/kg arimoclomol or the combination. Western blots of homogenates of cortex were probed with antibodies smc-100 (HSPA1A), SPA812 (inducible and constitutive HSP70), smc-151 (HSPA8), ADI-SPA-400 (HSP40), smc-149 (HSP90) or sc-1049 (HSPB1) and normalized to GAPDH or actin. Top row: Levels of HSPs were unchanged in cortex of FUS R521G compared to control mice. Rows 2–5: Levels were not affected by any drug treatments. Data are shown as mean ​± ​SD (3–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.

Article Snippet: Primary antibodies were: mouse anti-HSPA1A (StressMarq smc-100, 1:500), mouse anti-HSPA8 (StressMarq smc-151, 1:1000), rabbit anti-HSP70 (Enzo ADI-SPA812, 1:1000), goat anti-HSPB1 (Santa Cruz sc-1049, 1:1000), rabbit anti-HSP40 (Enzo ADI-SPA-400, 1:2000), mouse anti-HSP90 (StressMarq smc-149, 1:000), rabbit anti-α-tubulin (Abcam 15246, 1:10,000), mouse anti-tubulin acetylated (Sigma T6793, 1:10,000), mouse anti-STIP1(Abnova H00010963-M11, 1:1000), rat anti-HSF1 (StressMarq smc-118, 1:1000), rabbit anti-HDAC3 (Sigma H3034, 1:1000), mouse anti-α-tubulin (MediMabs MM-0163, 1:20,000), mouse anti-actin (Mp Biochemicals 691002, 1:20,000), mouse anti-GAPDH (MediMabs MM-0163, 1:5000), rabbit anti-SOD1 (Enzo ADI-SOD-101, 1:5000), rabbit anti-FUS ((Proteintech 11570-1-AP, 1:1000) and mouse anti-FUS (Santa Cruz, sc-47711, 1:1000).

Techniques: Expressing, Western Blot, Control, Two Tailed Test

No change in expression of heat shock proteins in gastrocnemius/soleus muscle of FUS R521G mice or with drug treatments. FUS R521G mice were treated for 28 days (7 days/week) i.p . beginning P60 with vehicle or the combination of 20 ​mg/kg arimoclomol and 10 ​mg/kg RGFP963. Homogenates of muscles were analyzed for levels of HSPA1A, HSPA8, HSP40, HSP90 or HSPB1 by Western blotting and normalized to GAPDH. Levels of HSPs were unchanged in muscles of FUS R521G compared to control mice (left column). Combination treatment had no effect on their levels (right column). Data are presented as mean ​± ​SD (7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.

Journal: Neurotherapeutics

Article Title: Reversal of cognitive deficits in FUS R521G amyotrophic lateral sclerosis mice by arimoclomol and a class I histone deacetylase inhibitor independent of heat shock protein induction

doi: 10.1016/j.neurot.2024.e00388

Figure Lengend Snippet: No change in expression of heat shock proteins in gastrocnemius/soleus muscle of FUS R521G mice or with drug treatments. FUS R521G mice were treated for 28 days (7 days/week) i.p . beginning P60 with vehicle or the combination of 20 ​mg/kg arimoclomol and 10 ​mg/kg RGFP963. Homogenates of muscles were analyzed for levels of HSPA1A, HSPA8, HSP40, HSP90 or HSPB1 by Western blotting and normalized to GAPDH. Levels of HSPs were unchanged in muscles of FUS R521G compared to control mice (left column). Combination treatment had no effect on their levels (right column). Data are presented as mean ​± ​SD (7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.

Article Snippet: Primary antibodies were: mouse anti-HSPA1A (StressMarq smc-100, 1:500), mouse anti-HSPA8 (StressMarq smc-151, 1:1000), rabbit anti-HSP70 (Enzo ADI-SPA812, 1:1000), goat anti-HSPB1 (Santa Cruz sc-1049, 1:1000), rabbit anti-HSP40 (Enzo ADI-SPA-400, 1:2000), mouse anti-HSP90 (StressMarq smc-149, 1:000), rabbit anti-α-tubulin (Abcam 15246, 1:10,000), mouse anti-tubulin acetylated (Sigma T6793, 1:10,000), mouse anti-STIP1(Abnova H00010963-M11, 1:1000), rat anti-HSF1 (StressMarq smc-118, 1:1000), rabbit anti-HDAC3 (Sigma H3034, 1:1000), mouse anti-α-tubulin (MediMabs MM-0163, 1:20,000), mouse anti-actin (Mp Biochemicals 691002, 1:20,000), mouse anti-GAPDH (MediMabs MM-0163, 1:5000), rabbit anti-SOD1 (Enzo ADI-SOD-101, 1:5000), rabbit anti-FUS ((Proteintech 11570-1-AP, 1:1000) and mouse anti-FUS (Santa Cruz, sc-47711, 1:1000).

Techniques: Expressing, Muscles, Western Blot, Control, Two Tailed Test

Drug treatments increased only HSPA8 in lumbar spinal cord (LSC) of endstage SOD1 G93A mice. B6 nontransgenic and SOD1 G93A transgenic mice were treated 5 days/week i.p. from P85 to endstage with vehicle, 20 ​mg/kg RGFP963, 20 ​mg/kg arimoclomol, or the combination. Homogenates of LSC were analyzed for levels of HSPA1A, HSPA8, HSP40 and HSP90 by Western blotting and normalized to GAPDH. (A) HSP levels were unchanged in LSC from SOD1 G93A mice compared to B6 control mice. (B–D) HSPA8 levels were significantly elevated by drug treatments, but HSPA1A, HSP40 and HSP90 remained unchanged. Data are shown as mean ​± ​SD (4 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.

Journal: Neurotherapeutics

Article Title: Reversal of cognitive deficits in FUS R521G amyotrophic lateral sclerosis mice by arimoclomol and a class I histone deacetylase inhibitor independent of heat shock protein induction

doi: 10.1016/j.neurot.2024.e00388

Figure Lengend Snippet: Drug treatments increased only HSPA8 in lumbar spinal cord (LSC) of endstage SOD1 G93A mice. B6 nontransgenic and SOD1 G93A transgenic mice were treated 5 days/week i.p. from P85 to endstage with vehicle, 20 ​mg/kg RGFP963, 20 ​mg/kg arimoclomol, or the combination. Homogenates of LSC were analyzed for levels of HSPA1A, HSPA8, HSP40 and HSP90 by Western blotting and normalized to GAPDH. (A) HSP levels were unchanged in LSC from SOD1 G93A mice compared to B6 control mice. (B–D) HSPA8 levels were significantly elevated by drug treatments, but HSPA1A, HSP40 and HSP90 remained unchanged. Data are shown as mean ​± ​SD (4 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.

Article Snippet: Primary antibodies were: mouse anti-HSPA1A (StressMarq smc-100, 1:500), mouse anti-HSPA8 (StressMarq smc-151, 1:1000), rabbit anti-HSP70 (Enzo ADI-SPA812, 1:1000), goat anti-HSPB1 (Santa Cruz sc-1049, 1:1000), rabbit anti-HSP40 (Enzo ADI-SPA-400, 1:2000), mouse anti-HSP90 (StressMarq smc-149, 1:000), rabbit anti-α-tubulin (Abcam 15246, 1:10,000), mouse anti-tubulin acetylated (Sigma T6793, 1:10,000), mouse anti-STIP1(Abnova H00010963-M11, 1:1000), rat anti-HSF1 (StressMarq smc-118, 1:1000), rabbit anti-HDAC3 (Sigma H3034, 1:1000), mouse anti-α-tubulin (MediMabs MM-0163, 1:20,000), mouse anti-actin (Mp Biochemicals 691002, 1:20,000), mouse anti-GAPDH (MediMabs MM-0163, 1:5000), rabbit anti-SOD1 (Enzo ADI-SOD-101, 1:5000), rabbit anti-FUS ((Proteintech 11570-1-AP, 1:1000) and mouse anti-FUS (Santa Cruz, sc-47711, 1:1000).

Techniques: Transgenic Assay, Western Blot, Control, Two Tailed Test

Increase in HSPA1A, HSPA8, HSP40, HSP90 and STIP1/HOP in gastrocnemius/soleus muscle of endstage SOD1 G93A mice, but no augmentation by drug treatments. B6 nontransgenic and SOD1 G93A mice were treated 5 days/week i.p. from P85 to endstage with vehicle, 20 ​mg/kg RGFP963, 20 ​mg/kg arimoclomol or the combination. Homogenates of gastrocnemius/soleus were analyzed for HSPA1A, HSPA8, HSP40, HSP90 or STIP1/HOP by Western blotting and normalized to GAPDH. (A) Levels of HSPA1A, HSPA8, HSP40, HSP90 and STIP1 were increased in muscle from SOD1 G93A mice compared to B6 control mice. (B-D) Combination drug treatment significantly reduced the elevation in HSP levels. Data are shown as mean ​± ​SD (5–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.

Journal: Neurotherapeutics

Article Title: Reversal of cognitive deficits in FUS R521G amyotrophic lateral sclerosis mice by arimoclomol and a class I histone deacetylase inhibitor independent of heat shock protein induction

doi: 10.1016/j.neurot.2024.e00388

Figure Lengend Snippet: Increase in HSPA1A, HSPA8, HSP40, HSP90 and STIP1/HOP in gastrocnemius/soleus muscle of endstage SOD1 G93A mice, but no augmentation by drug treatments. B6 nontransgenic and SOD1 G93A mice were treated 5 days/week i.p. from P85 to endstage with vehicle, 20 ​mg/kg RGFP963, 20 ​mg/kg arimoclomol or the combination. Homogenates of gastrocnemius/soleus were analyzed for HSPA1A, HSPA8, HSP40, HSP90 or STIP1/HOP by Western blotting and normalized to GAPDH. (A) Levels of HSPA1A, HSPA8, HSP40, HSP90 and STIP1 were increased in muscle from SOD1 G93A mice compared to B6 control mice. (B-D) Combination drug treatment significantly reduced the elevation in HSP levels. Data are shown as mean ​± ​SD (5–7 mice/group). Statistical analysis was by unpaired, two-tailed Student's t -test; ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001.

Article Snippet: Primary antibodies were: mouse anti-HSPA1A (StressMarq smc-100, 1:500), mouse anti-HSPA8 (StressMarq smc-151, 1:1000), rabbit anti-HSP70 (Enzo ADI-SPA812, 1:1000), goat anti-HSPB1 (Santa Cruz sc-1049, 1:1000), rabbit anti-HSP40 (Enzo ADI-SPA-400, 1:2000), mouse anti-HSP90 (StressMarq smc-149, 1:000), rabbit anti-α-tubulin (Abcam 15246, 1:10,000), mouse anti-tubulin acetylated (Sigma T6793, 1:10,000), mouse anti-STIP1(Abnova H00010963-M11, 1:1000), rat anti-HSF1 (StressMarq smc-118, 1:1000), rabbit anti-HDAC3 (Sigma H3034, 1:1000), mouse anti-α-tubulin (MediMabs MM-0163, 1:20,000), mouse anti-actin (Mp Biochemicals 691002, 1:20,000), mouse anti-GAPDH (MediMabs MM-0163, 1:5000), rabbit anti-SOD1 (Enzo ADI-SOD-101, 1:5000), rabbit anti-FUS ((Proteintech 11570-1-AP, 1:1000) and mouse anti-FUS (Santa Cruz, sc-47711, 1:1000).

Techniques: Western Blot, Control, Two Tailed Test

Quantitative expression profiles of UPP1, LACTB, and AHSA1 proteins in omics analysis. (A–C) Protein quantification values of UPP1, LACTB, and AHSA1 in PC vs para-PC group; (D–F) Protein quantification values of UPP1, LACTB, and AHSA1 in PD-PC vs MD-PC group. * P < 0.05. PD: Poorly differentiated; PC: Pancreatic cancer; MD: Moderately differentiated; UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1.

Journal: Biomolecules and Biomedicine

Article Title: UPP1 and AHSA1 as emerging biomarkers and targets in pancreatic cancer: A proteomic approach

doi: 10.17305/bb.2025.11958

Figure Lengend Snippet: Quantitative expression profiles of UPP1, LACTB, and AHSA1 proteins in omics analysis. (A–C) Protein quantification values of UPP1, LACTB, and AHSA1 in PC vs para-PC group; (D–F) Protein quantification values of UPP1, LACTB, and AHSA1 in PD-PC vs MD-PC group. * P < 0.05. PD: Poorly differentiated; PC: Pancreatic cancer; MD: Moderately differentiated; UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1.

Article Snippet: Sections were incubated overnight at 4 ∘ C with primary antibodies against UPP1 (1:100, Proteintech), AHSA1 (1:100, Boster), and LACTB (1:100, Boster).

Techniques: Expressing, Activity Assay

GEPIA database (A), RT-qPCR (B), and Western blot (C) detected the expression of UPP1, AHSA1 and LACTB in PC vs para-PC group. * P < 0.05, *** P < 0.001, **** P < 0.0001. PC: Pancreatic cancer; UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1; RT-qPCR: Reverse transcription-quantitative polymerase chain reaction; GEPIA: Gene Expression Profiling Interactive Analysis.

Journal: Biomolecules and Biomedicine

Article Title: UPP1 and AHSA1 as emerging biomarkers and targets in pancreatic cancer: A proteomic approach

doi: 10.17305/bb.2025.11958

Figure Lengend Snippet: GEPIA database (A), RT-qPCR (B), and Western blot (C) detected the expression of UPP1, AHSA1 and LACTB in PC vs para-PC group. * P < 0.05, *** P < 0.001, **** P < 0.0001. PC: Pancreatic cancer; UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1; RT-qPCR: Reverse transcription-quantitative polymerase chain reaction; GEPIA: Gene Expression Profiling Interactive Analysis.

Article Snippet: Sections were incubated overnight at 4 ∘ C with primary antibodies against UPP1 (1:100, Proteintech), AHSA1 (1:100, Boster), and LACTB (1:100, Boster).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Activity Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Gene Expression

H&E Staining (A), IF (B) and IHC (C) detected the expression of UPP1, AHSA1 and LACTB in PD-PC vs MD-PC group. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1; H&E: Hematoxylin and eosin; IF: Immunofluorescence; IHC: Immunohistochemistry; PD: Poorly differentiated; PC: Pancreatic cancer; MD: Moderately differentiated.

Journal: Biomolecules and Biomedicine

Article Title: UPP1 and AHSA1 as emerging biomarkers and targets in pancreatic cancer: A proteomic approach

doi: 10.17305/bb.2025.11958

Figure Lengend Snippet: H&E Staining (A), IF (B) and IHC (C) detected the expression of UPP1, AHSA1 and LACTB in PD-PC vs MD-PC group. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1; H&E: Hematoxylin and eosin; IF: Immunofluorescence; IHC: Immunohistochemistry; PD: Poorly differentiated; PC: Pancreatic cancer; MD: Moderately differentiated.

Article Snippet: Sections were incubated overnight at 4 ∘ C with primary antibodies against UPP1 (1:100, Proteintech), AHSA1 (1:100, Boster), and LACTB (1:100, Boster).

Techniques: Staining, Expressing, Activity Assay, Immunofluorescence, Immunohistochemistry

Kaplan–Meier survival analysis. UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1.

Journal: Biomolecules and Biomedicine

Article Title: UPP1 and AHSA1 as emerging biomarkers and targets in pancreatic cancer: A proteomic approach

doi: 10.17305/bb.2025.11958

Figure Lengend Snippet: Kaplan–Meier survival analysis. UPP1: Uridine Phosphorylase 1; LACTB: Lactamase Beta; AHSA1: Activator of HSP90 ATPase Activity 1.

Article Snippet: Sections were incubated overnight at 4 ∘ C with primary antibodies against UPP1 (1:100, Proteintech), AHSA1 (1:100, Boster), and LACTB (1:100, Boster).

Techniques: Activity Assay