Journal: BMC Genomics

Article Title: Benzo pyrene-induced DNA adducts and gene expression profiles in target and non-target organs for carcinogenesis in mice

doi: 10.1186/1471-2164-15-880

Figure Lengend Snippet: Genes found to be significantly differentially expressed (Welch t - test p < 0.05) in the same direction for all target organs or all non - target organs

Article Snippet: The assay codes were: GAPDH -4352339E, Trp53 -Mm00441964_g1, Pcna -Mm00448100_g1, Ccng1 -Mm00438084_m1, Ephx1 -Mm00468752_m1, Cyp1a1 -Mm00487218_m1, Cdh1 -Mm00486906_m1, Ctnnb1 -Mm01350394_m1, Apc -Mm00545877_m1, Stat3 -Mm00456961_m1, Erbb2 -Mm00658541_m1, Src -Mm00436783_m1, Hspa8 -Mm01731394_gH, Hspa5 -Mm00517691_m1, Fos -Mm00487425_m1, Myc -Mm00487804_m1, NFκB -Mm00476361_m1, Tnfα -Mm00443258_m1, Cav -Mm00483057_m1, Tubb5 -Mm00495804_m1, Cdkn2a -Mm00494449_m1, Ccnd2 -Mm00438071_m1.

Techniques: Control, Virus

Journal: BMC Genomics

Article Title: Benzo pyrene-induced DNA adducts and gene expression profiles in target and non-target organs for carcinogenesis in mice

doi: 10.1186/1471-2164-15-880

Figure Lengend Snippet: Gene expression changes validation with RT - PCR

Article Snippet: The assay codes were: GAPDH -4352339E, Trp53 -Mm00441964_g1, Pcna -Mm00448100_g1, Ccng1 -Mm00438084_m1, Ephx1 -Mm00468752_m1, Cyp1a1 -Mm00487218_m1, Cdh1 -Mm00486906_m1, Ctnnb1 -Mm01350394_m1, Apc -Mm00545877_m1, Stat3 -Mm00456961_m1, Erbb2 -Mm00658541_m1, Src -Mm00436783_m1, Hspa8 -Mm01731394_gH, Hspa5 -Mm00517691_m1, Fos -Mm00487425_m1, Myc -Mm00487804_m1, NFκB -Mm00476361_m1, Tnfα -Mm00443258_m1, Cav -Mm00483057_m1, Tubb5 -Mm00495804_m1, Cdkn2a -Mm00494449_m1, Ccnd2 -Mm00438071_m1.

Techniques: Gene Expression, Biomarker Discovery, Transformation Assay

Journal: BMC Genomics

Article Title: Benzo pyrene-induced DNA adducts and gene expression profiles in target and non-target organs for carcinogenesis in mice

doi: 10.1186/1471-2164-15-880

Figure Lengend Snippet: Genes significantly modulated by BaP in individual organs ( Welch ’ s t - test , p < 0.05 ) for at least three conditions

Article Snippet: The assay codes were: GAPDH -4352339E, Trp53 -Mm00441964_g1, Pcna -Mm00448100_g1, Ccng1 -Mm00438084_m1, Ephx1 -Mm00468752_m1, Cyp1a1 -Mm00487218_m1, Cdh1 -Mm00486906_m1, Ctnnb1 -Mm01350394_m1, Apc -Mm00545877_m1, Stat3 -Mm00456961_m1, Erbb2 -Mm00658541_m1, Src -Mm00436783_m1, Hspa8 -Mm01731394_gH, Hspa5 -Mm00517691_m1, Fos -Mm00487425_m1, Myc -Mm00487804_m1, NFκB -Mm00476361_m1, Tnfα -Mm00443258_m1, Cav -Mm00483057_m1, Tubb5 -Mm00495804_m1, Cdkn2a -Mm00494449_m1, Ccnd2 -Mm00438071_m1.

Techniques:

Journal: Redox Biology

Article Title: LACTB suppresses liver cancer progression through regulation of ferroptosis

doi: 10.1016/j.redox.2024.103270

Figure Lengend Snippet: HSPA8 is a target of LACTB. A. qRT-PCR testing the effects of LACTB overexpression and knockout on HSPA8 mRNA levels. B. Western blot testing the indicated protein levels after LACTB overexpression in SK-HEP-1 cells. C. Western blot testing the indicated protein levels in LACTB −/− cells reexpressing LACTB. D. Western blot testing the indicated protein levels in LACTB-expressing cells transfected with HSPA8-expressing plasmid. E. Western blot testing the indicated protein levels in LACTB −/− cells transfected with si-HSPA8. F–I. Detection of effects of LACTB overexpression or in combination with HSPA8 overexpression on Fe 2+ , lipid peroxidation, MDA levels and GSH/GSSG ratio in cells. Scale bar = 25 μm. J,K. Detection of effects of LACTB knockout or in combination with HSPA8 silencing on Fe 2+ and lipid peroxidation levels in HepG2 and SK-HEP-1 cells. Scale bar = 25 μm. L. Kaplan-Meier plotter database showing the overall survival rate of liver cancer patients with high or low HSPA8 expression. M. Xenograft tumour model testing the in vivo effects of LACTB or LACTB + HSPA8 on SK-HEP-1 cell growth. Scale bar = 1 cm. Student's t -test was used for A (left panel), one-way ANOVA with Tukey post-hoc test was used for A (right panel), F–K, M (right panel), and two-way ANOVA with Tukey post-hoc test was used for M (middle panel).

Article Snippet: The sections were then incubated with anti -LACTB (#18195-1-AP, Proteintech), anti -p53 (wild-type) (#MABE339, Sigma-Aldrich), and anti -HSPA8 (#M01024-1, BOSTER) primary antibodies and a DAKO peroxidase-labelled secondary antibody.

Techniques: Quantitative RT-PCR, Over Expression, Knock-Out, Western Blot, Expressing, Transfection, Plasmid Preparation, In Vivo

Journal: Redox Biology

Article Title: LACTB suppresses liver cancer progression through regulation of ferroptosis

doi: 10.1016/j.redox.2024.103270

Figure Lengend Snippet: LACTB drives ferroptosis via regulating the p53/HSPA8 axis. A. qRT-PCR testing HSPA8 mRNA levels in LACTB-expressing Huh7 and Hep3B cells. B, C. qRT-PCR and Western blot testing HSPA8 mRNA and protein levels in LACTB-expressing cells transfected with si-p53. D. Western blot testing p53 protein expression in LACTB-expressing SK-HEP-1 cells treated with 100 μg/mL cycloheximide for the indicated time. E. Western blot testing p53 ubiquitination levels in LACTB-expressing SK-HEP-1 cells. F. Western blot testing HSPA8 protein expression in LACTB-expressing p53 +/+ Hep3B cells. G. Luciferase reporter assay testing HSPA8 promoter activity in LACTB-expressing cells transfected with si-p53. H. p53 binding sites on HSPA8 promoter among different species. I. The schema showing the wild-type or mutant p53 binding motifs on HSPA8 or p21 promoter, and primer design positions for ChIP assay. J. ChIP assay testing the binding of p53 on the indicated regions of HSPA8 promoter. K. DNA pull-down assay using wild-type or mutant HSPA8 promoter probe, followed by Western blot analysis of p53 protein expression. L. Generation of SK-HEP-1 cells with endogenous mutation of p53 binding site in HSPA8 promoter using CRISPR/Cas9 gene editing technology with the indicated ssODN and gRNA. M. CCK-8 testing the viability of wild-type or mutant SK-HEP-1 cells treated with erastin and RSL3. N, O. Liperfluo and FerroOrange staining testing lipid peroxidation and Fe 2+ levels in wild-type or mutant SK-HEP-1 cells. Scale bar = 25 μm. P. Western blot testing HSPA8 protein levels in wild-type or mutant SK-HEP-1 cells with LACTB or p53 overexpression. Q. IHC staining of LACTB, wild-type (wt) p53 and HSPA8 in liver cancer tissue microarray, followed by analysis of their expression correlations. Scale bar = 50 μm. One-way ANOVA with Tukey post-hoc test was used for B and G. Student's t -test was used for J, N, O.

Article Snippet: The sections were then incubated with anti -LACTB (#18195-1-AP, Proteintech), anti -p53 (wild-type) (#MABE339, Sigma-Aldrich), and anti -HSPA8 (#M01024-1, BOSTER) primary antibodies and a DAKO peroxidase-labelled secondary antibody.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Transfection, Ubiquitin Proteomics, Luciferase, Reporter Assay, Activity Assay, Binding Assay, Mutagenesis, Pull Down Assay, CRISPR, CCK-8 Assay, Staining, Over Expression, Immunohistochemistry, Microarray

Journal: Redox Biology

Article Title: LACTB suppresses liver cancer progression through regulation of ferroptosis

doi: 10.1016/j.redox.2024.103270

Figure Lengend Snippet: LACTB potentiates the response of liver cancer to lenvatinib. A, B. FerroOrange and liperfluo staining testing Fe 2+ and lipid peroxidation levels in HepG2 and SK-HEP-1 cells treated with lenvatinib, respectively. Scale bar = 25 μm. C. Western blot testing LACTB protein expression in HepG2 and SK-HEP-1 cells treated with lenvatinib. D-G. FerroOrange, liperfluo staining, CCK-8 and colony formation assays respectively testing Fe 2+ , lipid peroxidation levels, viability and cloning ability in lenvatinib-treated cells with LACTB overexpression or knockout. H, I. Xenograft tumour model testing the in vivo effects of LACTB overexpression or knockout on the anti-tumour effect of lenvatinib. Scale bar = 1 cm. J. Western blot testing the indicated protein expression in the indicated groups. K. The proposed model showing that LACTB induced by lenvatinib promotes ferroptosis by increasing p53 protein stability and inhibiting HSPA8-mediated anti -ferroptosis pathway. One-way ANOVA with Tukey post-hoc test was used for A, B, D-G, I.

Article Snippet: The sections were then incubated with anti -LACTB (#18195-1-AP, Proteintech), anti -p53 (wild-type) (#MABE339, Sigma-Aldrich), and anti -HSPA8 (#M01024-1, BOSTER) primary antibodies and a DAKO peroxidase-labelled secondary antibody.

Techniques: Staining, Western Blot, Expressing, CCK-8 Assay, Cloning, Over Expression, Knock-Out, In Vivo

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

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: ChAT interacts with the HSP-associated E3 ubiquitin ligase C-terminus of HSC70-interaction protein (CHIP). (A) Immunoblots showing co-IP of ChAT with FLAG-CHIP from SN56 cells co-expressing either wild-type or mutant ChAT protein with FLAG-tagged CHIP. Control cells were transfected with either empty vector or to express either wild-type ChAT or FLAG-CHIP alone. (B) Co-IP of ChAT with FLAG-CHIP is enhanced for P17A/P19A- ( *** p ≤ 0.001), V18M- ( *** p ≤ 0.001), and A513T-ChAT ( * p ≤ 0.05) as compared to wild-type ChAT (one-way ANOVA with Dunnett's post-hoc test, mean ± SEM, n = 5). (C) Co-IP of wild-type and mutant ChAT with endogenous CHIP following anti-ChAT co-IP from ChAT-expressing SN56 cells ( n = 3). (D) Detection of in situ ChAT interactions with endogenous CHIP by proximity ligation assay (PLA) in SN56 cells expressing wild-type ChAT. Formalin-fixed cells were first co-labeled with goat anti-ChAT together with rabbit anti-CHIP 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-CHIP interactions were imaged by confocal microscopy. Positive in situ ChAT-CHIP interactions were visualized as fluorescent red dots while nuclei were stained with DAPI (blue). Control cells were either transfected with empty vector or primary antibodies omitted from the assay (No 1° antibodies). Images are representative of 4 independent experiments; scale bars are 10 μm. (E) siRNA-mediated knock-down of CHIP has no effect on the steady-state protein levels of either wild-type or mutant ChAT. ChAT-expressing SN56 cells were co-transfected with 25 nM of either anti-CHIP siRNA or scramble-control siRNA for 72 h. Control cells were mock-transfected ( 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: Western Blot, Co-Immunoprecipitation Assay, Expressing, Mutagenesis, Transfection, Plasmid Preparation, In Situ, Proximity Ligation Assay, Labeling, Incubation, DNA Ligation, Amplification, Confocal Microscopy, Staining

Journal: bioRxiv

Article Title: Chaperone-mediated autophagy is an overlooked pathway for mutant α1-antitrypsin Z degradation

doi: 10.1101/2023.11.24.568525

Figure Lengend Snippet: A 、 C:HEK 293 stably expressing GFP-ATZ were transfected with an empty vector and LAMP2A or HSC70 for 48 hours, cells were lysed and subjected to immunoprecipitation with antibodies against IgG or Flag. Aliquots of IP proteins and input proteins were analyzed by immunoblot. B: HEK 293 stably expressing GFP-ATZ (green) were transiently transfected with HSC70 or LAMP2A for 48 h. Immunofluorescence microscopic images of GFP-ATZ (green) and LAMP2A(red) in HEK 293 stably expressing GFP-ATZ cells were obtained after staining with anti-LAMP2A antibodies. Each nucleus was stained with DAPI (blue). The co-localization of LAMP2A and ATZ-GFP were visualized as yellow color caused by superimposing of red and green. Plots of pixel intensity along the white line rows of images to the left of each plot, colors as in merged images. Scale bars: 10 μm D: GFP-ATZ-expressing HEK 293 cells were transfected with an empty vector or a vector encoding LAMP2A for 36 hours. Subsequently, they were treated with CQ (10μM) for an additional duration of 4, 5, or 6 hours. Following the treatment, the cells were lysed and underwent western blot analysis. E: HEK 293 cells expressing GFP-ATZ were treated with DMSO or QX77 (10μM) for 48 hours. Cell lysates were immunoblotted with the indicated antibodies. F: Schematic depiction of a full-length LAMP2A construct and LAMP2A-mutant construct. TS: Transmembrane structure. G: GFP-ATZ-expressing HEK 293 cells were transfected with full-length LAMP2A, LAMP2A-mutant, or LAMP2A-truncate constructs for 48 hours. The protein levels of GFP-ATZ in different transfections were quantitatively analyzed. TR: LAMP2A truncate, Mu: LAMP2A mutant. H: HEK 293 cells were transfected with LAMP2A, LAMP2A-mutant, or LAMP2A-truncate along with FLAG-ATZ for 24 hours. The cell lysates were then subjected to immunoprecipitation (IP) using an anti-Flag antibody, and the precipitates were analyzed by western blotting. TR: LAMP2A truncate, MU: LAMP2A mutant. Data were presented as the mean ± SEM of 3 or more independent experiments. p <0.05.*; p <0.01.**; p <0.001.***; p <0.0001.****; NC, no significance. vs. indicated group.

Article Snippet: LAMP2A and HSC70 were cloned into pCDH-CMV (72265, Addgene) by PCR amplifying the ORFs from cDNA templates of HSC70 (19514, Addgene) and LAMP2A (86146, Addgene), followed by digestion with BamHI and EcoRI restriction enzymes.

Techniques: Stable Transfection, Expressing, Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Immunofluorescence, Staining, Construct, Mutagenesis

Journal: bioRxiv

Article Title: Chaperone-mediated autophagy is an overlooked pathway for mutant α1-antitrypsin Z degradation

doi: 10.1101/2023.11.24.568525

Figure Lengend Snippet: A: The cartoon illustrates the structure of alpha-1 antitrypsin, with the pentapeptide sequence (121QELLR125) highlighted in orange. The combined ribbon representation and stick model showing the overall structure and the CMA motif of ATZ protein. This model was generated using PyMOLTM based on the Protein Data Bank code 3CWM. B 、 C:HEK 293 cells transfected with LAMP2A or HSC70 and ATZ-FLAG or AA-ATZ-FLAG for 48h. Lysates were immunoprecipitated (IP) with either IgG, anti-LAMP2A antibody and probed for ATZ-FLAG, AA-ATZ-FLAG and LAMP2A or HSC70. D: HEK 293 cells were con-transfected for 48 hours with LAMP2A expressing vectors or vector coding wide type ATZ or AA-ATZ mutant. Representative confocal images showing the colocalization of ATZ-FLAG (red) or AA-ATZ-FLAG (red) and LAMP2A(green) in HEK 293T cells. Scale bars: 10 μm E: HEK 293 cells were co-transfected with LAMP2A expressing vectors or vectors encoding wild-type ATZ or AA-ATZ mutant for 48 hours. The cells were lysed, and western blot analysis was conducted to assess the expression of GFP-ATZ. F: HEK 293 stably expressing LAMP2A cells were transfected with wide type ATZ or AA-ATZ expressing vectors. The transfected cells were cultured for 24 h before being further incubated with cyclohexamide (CHX;10 μg/ml) for the indicated time. The levels of A1AT at different time points were detected by Western blot.

Article Snippet: LAMP2A and HSC70 were cloned into pCDH-CMV (72265, Addgene) by PCR amplifying the ORFs from cDNA templates of HSC70 (19514, Addgene) and LAMP2A (86146, Addgene), followed by digestion with BamHI and EcoRI restriction enzymes.

Techniques: Sequencing, Generated, Transfection, Immunoprecipitation, Expressing, Plasmid Preparation, Mutagenesis, Western Blot, Stable Transfection, Cell Culture, Incubation

Journal: bioRxiv

Article Title: Chaperone-mediated autophagy is an overlooked pathway for mutant α1-antitrypsin Z degradation

doi: 10.1101/2023.11.24.568525

Figure Lengend Snippet: A: HEK 293 cells expressing GFP-ATZ were transfected with LAMP2A or HSC70 for 48 hours. Cell viability analysis was performed using the CellTiter-Glo luminescent cell viability assay. B: TMRE signals in HEK 293 cells expressing GFP-ATZ were detected by fluorescence microscopy. Scale bars: 50 μm. CCCP: negative control. Ctrl: pCDH empty plasmid. C: HEK 293 cells expressing GFP-ATZ were transfected with either LAMP2A or HSC70, then stained with TMRE or CCCP for flow cytometry analysis. CCCP: negative control. Ctrl: pCDH empty plasmid. D: A volcano plot was generated to compare the log2 fold change (logFC) versus the p-value between control cells and LAMP2A-expressing cells. Grey circles represent RNAs with a p-value > 0.05, blue circles represent downregulated RNAs with a p-value < 0.05 and fold change greater than 2, while red circles represent upregulated RNAs with a p-value < 0.05 and fold change greater than 2, n=2. E: The expression levels of 16 genes displaying downregulation in HEK 293-GFP-ATZ cells under LAMP2A transfection conditions were subjected to Gene Ontology (GO) functional enrichment analysis (Biological Process). F: The corresponding heatmap highlights the disparity in RNA expression levels related to the heat shock protein family between the overexpression of the LAMP2A plasmid and the empty vector in GFP-ATZ-expressing HEK 293 cells.

Article Snippet: LAMP2A and HSC70 were cloned into pCDH-CMV (72265, Addgene) by PCR amplifying the ORFs from cDNA templates of HSC70 (19514, Addgene) and LAMP2A (86146, Addgene), followed by digestion with BamHI and EcoRI restriction enzymes.

Techniques: Expressing, Transfection, Cell Viability Assay, Fluorescence, Microscopy, Negative Control, Plasmid Preparation, Staining, Flow Cytometry, Generated, Control, Functional Assay, RNA Expression, Over Expression

Journal: bioRxiv

Article Title: Chaperone-mediated autophagy is an overlooked pathway for mutant α1-antitrypsin Z degradation

doi: 10.1101/2023.11.24.568525

Figure Lengend Snippet: Besides proteasome and autophagy pathways, CMA also plays a role in alpha-1 antitrypsin Z (ATZ) degradation. Specifically, the heat shock cognate 70 (HSC70) protein recognizes the ATZ protein through the pentapeptide sequence 121QELLR125 and transports it to lysosome-associated membrane protein 2A (LAMP2A) on the lysosomal membrane. Augmenting CMA activity through LAMP2A overexpression or AR7 utilization enhances ATZ digestion via this pathway, thereby alleviating cellular stress, including ER stress and mitochondrial impairment.

Article Snippet: LAMP2A and HSC70 were cloned into pCDH-CMV (72265, Addgene) by PCR amplifying the ORFs from cDNA templates of HSC70 (19514, Addgene) and LAMP2A (86146, Addgene), followed by digestion with BamHI and EcoRI restriction enzymes.

Techniques: Sequencing, Membrane, Activity Assay, Over Expression

Journal: Cell reports

Article Title: Chronic hypoxia stabilizes 3βHSD1 via autophagy suppression

doi: 10.1016/j.celrep.2023.113575

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Plasmid pCMV3-HSPA8-Myc expressing C-terminal Myc tagged HSPA8 , Sino Biological , HG11329-CM.

Techniques: Recombinant, Transfection, Mutagenesis, Viability Assay, Sequencing, Negative Control, shRNA, Plasmid Preparation, Expressing, Variant Assay, Software