Journal: Journal of Biomedical Science

Article Title: Degradation of neurodegenerative disease-associated TDP-43 aggregates and oligomers via a proteolysis-targeting chimera

doi: 10.1186/s12929-023-00921-7

Figure Lengend Snippet: Examination of the C-TDP-43 disaggregation and beneficial effects of PROTACs 1–4 . A Representative images of eGFP-C-TDP-43-expressing Neuro-2a cells with or without PROTAC 1 – 4 (5 μM). Scale bar = 10 μm. B Filter trap assay of eGFP-C-TDP-43 expressed Neuro-2a cells in the presence and absence of PROTACs 1 – 4 (5 μM). The cell lysate was either loaded on cellulose acetate (CA) or nitrocellulose (NC) membrane probed with TDP-43 (C-terminal) antibody and β-actin antibody (loading control), respectively. C Quantification of blots in panel B. D AlamarBlue reduction assay of eGFP-C-TDP-43 expressed Neuro-2a cells treated with PROTACs 1 – 4 (5 μM). E Western blot of eGFP-C-TDP-43 transfected Neuro-2a cells treated with various concentrations of PROTAC 2 . The RIPA-insoluble fraction and RIPA-soluble fraction of Neuro-2a lysate were further probed with GFP and GAPDH antibody, respectively. All the statistic results were quantified by ImageJ and shown as mean ± SD (n ≥ 3). Data were analyzed by one-way ANOVA with Dunnett post-hoc test (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Blots were blocked with 5% bovine serum albumin (BSA, Sigma) in 0.1% PBST for at least 1 h. After blocking, blots were subjected to incubation with the primary antibodies TDP-43 (C-terminal) (1:1000, Proteintech, 12,892-1-AP), TDP-43 (1:1000, Abcam, ab104223), p-TDP-43 (pS409/410) (1:1000, Cosmo Bio, TIP-PTD-M01), GFP (1:1000, Abcam, ab183734), A11 (1:1000, Invitrogen, AHB0052), GAPDH (1:10,000, GeneTex, GTX627408), GSPT1 (1:1000, Proteintech, 10,763-1-AP), flag M2 (1:1000, Sigma, F1804), LC3B (1:1000, cell signaling, #2775), γH2A.X (phosphor-Ser139) (1:1000, Merck Millipore, 05–636), HSP70 (1:1000, Proteintech, 10,995-1-AP), HMGB1 (1:1000, Abcam, ab18256) or β-actin (1:10,000, GeneTex, GTX109639) in 2–5% BSA and incubated overnight at 4 °C on a shaker.

Techniques: Expressing, TRAP Assay, Membrane, Control, Western Blot, Transfection

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Antibody responses to galectin-8, TARP and TRAP1 in prostate cancer patients treated with a GM-CSF-secreting cellular immunotherapy

doi: 10.1007/s00262-010-0858-5

Figure Lengend Snippet: Anti-human galectin-8, TARP and TRAP1 antibodies in patients treated with GVAX immunotherapy for prostate cancer. A longitudinal analysis of humoral reactivity to human galectin-8 (upper panels), TARP (middle panels) and TRAP1 (lower panels) in two G-0010 patients: 057 (a) and 202 (b). Serum samples at representative time points over the course of immunotherapy treatment were diluted 1:100 and incubated with 200 ng of the target antigen in an ELISA assay. An anti-human pan-IgG secondary antibody was used for detection. Arrows denote immunization with irradiated GM-CSF-secreting tumor cells

Article Snippet: Full-length LGALS8 , TARP and TRAP1 cDNAs were obtained from Origene (Rockville, MD, USA), PCR cloned with a C-terminus Flag-tag and transfected into 293 cells for protein production.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Irradiation

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Antibody responses to galectin-8, TARP and TRAP1 in prostate cancer patients treated with a GM-CSF-secreting cellular immunotherapy

doi: 10.1007/s00262-010-0858-5

Figure Lengend Snippet: RNA encoding the TARP and TRAP1 antigens shows enhanced expression in prostate cancer. RNA was extracted from normal (N = 8) and cancerous (N = 40) prostate tissue samples, cDNA was synthesized and then normalized for ACTB expression. Q-PCR primer and probe sets specific for LGALS8 (a), TARP (b), TRAP1 (c) and ACTB (d; control) were used to determine RNA transcript levels. C t denotes cycle threshold. A decrease in C t represents an increase in RNA transcript

Article Snippet: Full-length LGALS8 , TARP and TRAP1 cDNAs were obtained from Origene (Rockville, MD, USA), PCR cloned with a C-terminus Flag-tag and transfected into 293 cells for protein production.

Techniques: Expressing, Synthesized, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Antibody responses to galectin-8, TARP and TRAP1 in prostate cancer patients treated with a GM-CSF-secreting cellular immunotherapy

doi: 10.1007/s00262-010-0858-5

Figure Lengend Snippet: Kaplan–Meier estimates of overall survival in G-9803/G-0010 mCRPC patients with (solid lines) or without (dashed lines) an induced antibody response to galectin-8 (a), TARP (b) or TRAP1 (c)

Article Snippet: Full-length LGALS8 , TARP and TRAP1 cDNAs were obtained from Origene (Rockville, MD, USA), PCR cloned with a C-terminus Flag-tag and transfected into 293 cells for protein production.

Techniques:

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Antibody responses to galectin-8, TARP and TRAP1 in prostate cancer patients treated with a GM-CSF-secreting cellular immunotherapy

doi: 10.1007/s00262-010-0858-5

Figure Lengend Snippet: Humoral responses to TARP are observed in CRPC patients before immunotherapy treatment. Comparison of antibody reactivity (OD at 450 nm) between normal (N = 25) and CRPC patients before the initiation of immunotherapy treatment (N = 60) to the galectin-8 (a), TARP (b) and TRAP1 (c) antigens. Serum samples were diluted 1:100 and incubated with 200 ng of the target antigen in an ELISA assay using an anti-human pan-IgG secondary antibody for detection

Article Snippet: Full-length LGALS8 , TARP and TRAP1 cDNAs were obtained from Origene (Rockville, MD, USA), PCR cloned with a C-terminus Flag-tag and transfected into 293 cells for protein production.

Techniques: Comparison, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Antibody responses to galectin-8, TARP and TRAP1 in prostate cancer patients treated with a GM-CSF-secreting cellular immunotherapy

doi: 10.1007/s00262-010-0858-5

Figure Lengend Snippet: TARP and TRAP1 protein is over-expressed in prostate cancer. Representative photomicrographs of normal prostate (a, c) and prostate adenocarcinoma (b, d) tissue stained with an anti-TARP (upper panels) or anti-TRAP1 (lower panels) antibody. Original magnifications: ×5 (main-panel) and ×40 (sub-panel). Black boxed area (×5 magnification) indicates the area examined under higher magnification (×40) presented below. e, f TARP and TRAP1 RNA expression in a panel of cell lines as evaluated by quantitative PCR. RNA was extracted from PC-3, LNCaP, 293, K-562 and HeLa cells, cDNA synthesized and expression determined using gene specific Q-PCR primer and probe sets. C t denotes cycle threshold. A decrease in C t represents an increase in RNA transcript

Article Snippet: Full-length LGALS8 , TARP and TRAP1 cDNAs were obtained from Origene (Rockville, MD, USA), PCR cloned with a C-terminus Flag-tag and transfected into 293 cells for protein production.

Techniques: Staining, RNA Expression, Real-time Polymerase Chain Reaction, Synthesized, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Antibody responses to galectin-8, TARP and TRAP1 in prostate cancer patients treated with a GM-CSF-secreting cellular immunotherapy

doi: 10.1007/s00262-010-0858-5

Figure Lengend Snippet: Target antigens in immunotherapy treated patients ( N = 13) identified by autoantibody profiling of patients serum using protein microarray analysis

Article Snippet: Full-length LGALS8 , TARP and TRAP1 cDNAs were obtained from Origene (Rockville, MD, USA), PCR cloned with a C-terminus Flag-tag and transfected into 293 cells for protein production.

Techniques: Microarray, Binding Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA SNHG14 promotes hepatocellular carcinoma progression by regulating miR-876-5p/SSR2 axis

doi: 10.1186/s13046-021-01838-5

Figure Lengend Snippet: SSR2 is a downstream target of miR-876-5p. a The predicted results of miR-876-5p downstream target genes from four databases. b-c Relative expression of the nine downstream target genes when stimulated with miR-876-5p mimic or miR-876-5p inhibitor through RT-qPCR analysis. d Comparison the expression of SSR2 in carcinoma and adjacent tissues in TCGA-LIHC. e Comparison the expression of SSR2 in different grades (G1/G2 vs G3/G4) in TCGA-LIHC. f The correlation of the expression of SNHG14 and SSR2 in TCGA-LIHC. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: SSR2 antibody was purchased from proteintech (#10278–1-AP; Wuhan, Hubei, China).

Techniques: Expressing, Quantitative RT-PCR, Comparison

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Long noncoding RNA SNHG14 promotes hepatocellular carcinoma progression by regulating miR-876-5p/SSR2 axis

doi: 10.1186/s13046-021-01838-5

Figure Lengend Snippet: SNHG14 promotes HCC progression by regulating miR-876-5p/SSR2 axis. a-b The CCK8 assays in HLF and Hep3B with co-transfection of siSNHG14 and miR-876-5p inhibitor or pcDNA3.1/SSR2. c-d The transwell assays in HLF and Hep3B with co-transfection of siSNHG14 and miR-876-5p inhibitor or pcDNA3.1/SSR2. Scale bars: 100x = 100 um. e Images of tumors dissected from nude mice that were transplanted with SNHG14 knockdown HLF cells, SNHG14 knockdown plus SSR2 overexpression HLF cells and its negative control HLF cells. And tumor volume and tumor weight in these two groups were quantified. f The number of lung metastatic nodules in these three groups were quantified. Scale bars: 40x = 250 um; 100x = 100 um. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: SSR2 antibody was purchased from proteintech (#10278–1-AP; Wuhan, Hubei, China).

Techniques: Cotransfection, Knockdown, Over Expression, Negative Control