Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Conditions and titrations of the 10 antibodies

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques: Control

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Expressions of the 10 antibodies in 94 cases of leiomyoma, ESS and ULMS

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques:

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Positive expression of ASC1 in (A) low grade (LG) endometrial stromal sarcoma (ESS), (B) ‘LG’ uterine leiomyosarcoma (ULMS), (C) leiomyoma (LM). ASC1 was strongly expressed in cytoplasmic and nuclear patterns. Positive expression of stathmin1 in (D) in ESS, (E) leiomyosarcoma and (F) LM. Stathmin1 was strongly expressed in cytoplasmic and nuclear patterns.

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques: Expressing

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Comparison of biomarker expressions between all ESS and ULMS

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques: Comparison, Biomarker Discovery

Journal: Journal of clinical pathology

Article Title: Immunohistochemical panel to differentiate endometrial stromal sarcoma, uterine leiomyosarcoma and leiomyoma: something old and something new

doi: 10.1136/jclinpath-2015-202915

Figure Lengend Snippet: Comparison of biomarker expressions between LG ESS and ‘LG’ ULMS

Article Snippet: ASC1 , Novus , 1:100 , PH 8.0 20 min , liver.

Techniques: Comparison, Biomarker Discovery

Journal: bioRxiv

Article Title: The Ski2 helicase ASCC3 unwinds DNA upon fork stalling to control replication stress responses

doi: 10.1101/2025.07.24.666583

Figure Lengend Snippet: ASCC3 unwinds DNA to promote ssDNA-RPA accumulation upon replication stress. ( A ) Quantification of the percentage of cells with ≥ 10 BrdU foci in U2OS WT and ASCC3-KO cells that were pulse-labeled with BrdU for 20 min prior to their treatment with or without 4 mM HU for 4 h. A total of 802-825 cells per condition were scored in blind. SDs from three independent experiments are indicated in this and subsequent panels. *** P <0.001. ( B ) Quantification of the percentage of cells with ≥ 30 BrdU foci in U2OS WT and ASCC3-KO cells that were labeled with BrdU for 20 h prior to their treatment with or without 4 mM HU for 4 h. A total of 800-849 cells per condition were scored in blind. *** P <0.001. ( C ) Quantification of the percentage of cells with ≥ 10 BrdU foci in U2OS ASCC3-KO cells expressing various Myc-ASCC3 alleles as indicated. These cells were pulse-labeled with BrdU for 20 min prior to their treatment with HU. A total of 601-641 cells per condition were scored in blind. *** P <0.001. ( D ) Quantification of the percentage of cells with ≥ 30 BrdU foci in U2OS ASCC3-KO cells expressing various Myc-ASCC3 alleles as indicated. These cells were labeled with BrdU for 20 h prior to their treatment with HU. A total of 502-521 cells per condition were scored in blind. * P <0.05; ** P <0.01. ( E ) Quantification of EdU+ cells with ≥ 30 RPA32 foci in U2OS WT and ASCC3-KO cells that were pulse-labeled with EdU for 10 min prior to treatment with or without 4 mM HU for 6 h. A total of 800-873 cells per condition were scored in blind. *** P <0.001. ( F ) Quantification of the percentage of EdU+ cells with ≥ 30 RPA32 foci in U2OS ASCC3-KO cells expressing various Myc-ASCC3 alleles as indicated. A total of 801-856 cells per condition were scored in blind. *** P <0.001. ( G ) Quantification of the percentage of cells with ≥ 10 BrdU foci in U2OS WT and TRIP4-KO cells that were pulse-labeled with BrdU for 20 min prior to their treatment with or without HU. A total of 601-650 cells per condition were scored in blind. ** P <0.01. ( H ) Quantification of the percentage of EdU+ cells with ≥ 30 RPA32 foci in U2OS WT and TRIP4-KO cells that were pulse-labeled with EdU for 10 min prior to their treatment with or without HU. A total of 742-977 cells per condition were scored in blind. ** P <0.01. ( I ) Schematic diagram of in vitro RPA binding assays. ( J ) Western blot analyses of recombinant RPA70 recovered from streptavidin pulldown of biotinylated ssDNA from indicated reactions. Immunoblotting was done with an anti-RPA70 antibody. ( K ) Quantification of recombinant RPA70 from (J). * P <0.05.

Article Snippet: Antibodies used include: ASCC2 (1:1000; A304-020A, Bethyl Laboratories); ASCC2 (1:5000; 1152-1-AP, Proteintech); ASCC3 (1:400; 17627-1-AP, Proteintech); ASCC3 (1:1000; PA5-56794, Invitrogen); Biotin (1:100000; A150-109A, Bethyl Laboratory); Biotin (1:100000; 200-002-211, Jackson ImmunoResearch); BrdU (1:100; 347580, BD Biosciences); BrdU (1:500; MAB3222, Millipore); BrdU (BU1/75 [ICR1]) (1:800; NB500-169, Novus Biologicals); 53BP1 (1:2000; 612522, BD Biosciences); BRCA1 (1:5000; 07-434, Millipore); BRCA2 (1:2000; 29450-1-AP, Proteintech); CHK1 (1:250; sc-7898, Santa cruz); CHK1-pS345 (1:1000; 2348S, Cell Signaling); CSB (1:200; ab66598, Abcam); FASN (1:20000; 10624-2-AP, Proteintech); GFP (1:1000; 50430-2-AP, Proteintech); HA (1:500; #2367, Cell Signaling); HLTF (1:1000; A300-229A, Bethyl Laboratory); Myc (1:1000; 9E10, Calbiochem); PCNA (1:2000; sc-56, Santa Cruz); PCNA (1:600 for PLA or 1:4000 for western blot; 10205-2-AP, Proteintech); Ubiquityl-PCNA (K164) (1:1000; 13439T, Cell Signaling); PRIMPOL (1:1000; 29824-1-AP, Proteintech); RAD51 (1:2000, ab63801, Abcam); RFWD3 (1:1000; 19893-1-AP, Proteintech); RPA32 (1:10000; NB100-332, Novus Biologicals); RPA32 (1:200; ab2175, Abcam); RPA32-pS33 (1:50000; A300-246A, Bethyl Laboratories); RPA70 (1:2000; 2267S, Cell Signaling); SHPRH (21995-1-AP, Proteintech); SMARCAL1 (1:100, sc-166209, Santa cruz); SMARCAL1 (1:2000; GTX109468, GenTex); TRIP4 (1:1000; 12324-1-AP, Proteintech); α-tubulin (1:20,000; T9026, Sigma); γ-tubulin (1:20,000; GTU88, Sigma).

Techniques: Labeling, Expressing, In Vitro, Binding Assay, Western Blot, Recombinant

Journal: bioRxiv

Article Title: The Ski2 helicase ASCC3 unwinds DNA upon fork stalling to control replication stress responses

doi: 10.1101/2025.07.24.666583

Figure Lengend Snippet: ASCC3 antagonizes RAD51-mediated recombination. ( A ) Quantification of the CldU/IdU ratio in HU-treated U2OS WT and ASCC3-KO cells expressing control siRNA or siRNA against PRIMPOL (siPRIMPOL). DNA fiber experiments were performed twice independently with reproducible data in this, 6B, and 6C panels. Data from one representative experiment are shown as scatter plot graphs with the mean indicated in this, 6B, and 6C panels. A total of 407-421 fibers per condition were analyzed. The P -value was determined using a non-parametric Mann-Whitney rank-sum t -test in this and subsequent panels. **** P <0.0001. ( B ) Quantification of the CldU/IdU ratio in U2OS WT and ASCC3-KO cells that were treated with HU in the presence or absence of REV1 inhibitor JH-RE-06 (REV1i). A total of 413-414 fibers per condition were analyzed. **** P <0.0001. ( C ) Quantification of the CldU/IdU ratio in HU-treated U2OS WT and ASCC3-KO cells expressing control siRNA or two independent siRNA against RAD51 as indicated. A total of 407-415 fibers per condition were analyzed. **** P <0.0001. ( D ) Quantification of EdU+ cells with ≥ 20 RAD51 foci in U2OS WT and ASCC3-KO cells that were pulse-labeled with EdU for 10 min prior to treatment with or without 4 mM HU for 6 h. A total of 801-853 cells per condition were scored in blind. SDs from three independent experiments are indicated in this, and 6E-6G panels. *** P <0.001. ( E ) Quantification of EdU+ cells with ≥ 20 RAD51 foci in U2OS ASCC3-KO cells expressing various Myc-ASCC3 alleles as indicated. A total of 802-827 cells per condition were scored in blind. ** P <0.01; *** P <0.001. ( F ) Quantification of cells with ≥ 10 IR-induced RAD51 foci in U2OS WT and ASCC3-KO cells that were treated with 10 Gy IR prior to fixation. A total of 503-554 cells per condition were scored in blind. ** P <0.01. ( G ) Quantification of the percentage of cells with restoration of GFP expression following HR-mediated repair of I-SceI-induced DSBs. ** P <0.01. ( H ) Schematic diagram of strand exchange assays. ( I ) In vitro strand exchange assays. Top panel: Recombinant RAD51 (270 nM) was premixed with 1µM of 3’-ssDNA tail for 5 min, followed by further incubation with 350 nM of labeled duplex DNA and increasing concentration (150 nM, 300 nM, 600 nM) of ASCC3 HR WT-TRIP4 or ASCC3 HR -DD-AA-TRIP4 for 15 min. * indicates the 32 P-labeled DNA end. Bottom panel: Quantification of strand exchange efficiency (%) relative to RAD51 alone reaction from top panel. SDs from four independent experiments are shown. * P <0.05; **** P <0.0001.

Article Snippet: Antibodies used include: ASCC2 (1:1000; A304-020A, Bethyl Laboratories); ASCC2 (1:5000; 1152-1-AP, Proteintech); ASCC3 (1:400; 17627-1-AP, Proteintech); ASCC3 (1:1000; PA5-56794, Invitrogen); Biotin (1:100000; A150-109A, Bethyl Laboratory); Biotin (1:100000; 200-002-211, Jackson ImmunoResearch); BrdU (1:100; 347580, BD Biosciences); BrdU (1:500; MAB3222, Millipore); BrdU (BU1/75 [ICR1]) (1:800; NB500-169, Novus Biologicals); 53BP1 (1:2000; 612522, BD Biosciences); BRCA1 (1:5000; 07-434, Millipore); BRCA2 (1:2000; 29450-1-AP, Proteintech); CHK1 (1:250; sc-7898, Santa cruz); CHK1-pS345 (1:1000; 2348S, Cell Signaling); CSB (1:200; ab66598, Abcam); FASN (1:20000; 10624-2-AP, Proteintech); GFP (1:1000; 50430-2-AP, Proteintech); HA (1:500; #2367, Cell Signaling); HLTF (1:1000; A300-229A, Bethyl Laboratory); Myc (1:1000; 9E10, Calbiochem); PCNA (1:2000; sc-56, Santa Cruz); PCNA (1:600 for PLA or 1:4000 for western blot; 10205-2-AP, Proteintech); Ubiquityl-PCNA (K164) (1:1000; 13439T, Cell Signaling); PRIMPOL (1:1000; 29824-1-AP, Proteintech); RAD51 (1:2000, ab63801, Abcam); RFWD3 (1:1000; 19893-1-AP, Proteintech); RPA32 (1:10000; NB100-332, Novus Biologicals); RPA32 (1:200; ab2175, Abcam); RPA32-pS33 (1:50000; A300-246A, Bethyl Laboratories); RPA70 (1:2000; 2267S, Cell Signaling); SHPRH (21995-1-AP, Proteintech); SMARCAL1 (1:100, sc-166209, Santa cruz); SMARCAL1 (1:2000; GTX109468, GenTex); TRIP4 (1:1000; 12324-1-AP, Proteintech); α-tubulin (1:20,000; T9026, Sigma); γ-tubulin (1:20,000; GTU88, Sigma).

Techniques: Expressing, Control, MANN-WHITNEY, Labeling, In Vitro, Recombinant, Incubation, Concentration Assay

Journal: Cancers

Article Title: N-Acetylcysteine Promotes Metastatic Spread of Melanoma in Mice

doi: 10.3390/cancers14153614

Figure Lengend Snippet: Effect of inhibition of Cys/cystine uptake or GSH synthesis on GSH levels and metastatic activity in B16-F10 cells. Cancer cells were transfected in vitro, as explained under Material and Methods, before their in vivo inoculation. B16-F10 cells were inoculated i.v. (portal vein). NAC (240 mg/kg) was administered orally for 10 days, starting 120 min after B16-F10 cells inoculation. All measurements were performed in the liver or in metastatic cells isolated from the liver 10 days after tumor inoculation. Results obtained using melanoma cells treated with scrambled RNA sequences were not significantly different from those displayed as non-treated controls. * Significantly different p < 0.01, comparing shRNA-treated cells versus controls; + Significantly different p < 0.01, comparing NAC-treated mice versus mice treated with vehicle (n = 9–10).

Article Snippet: Knockdown of SLC7A10 (codes the ASC1 Na + -independent neutral amino acid transporter) was performed using an ASC1 shRNA Plasmid sc-39159-SH from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Inhibition, Activity Assay, Transfection, In Vitro, In Vivo, Isolation, shRNA, Control

Journal: Nature communications

Article Title: Extended DNA threading through a dual-engine motor module of the activating signal co-integrator 1 complex.

doi: 10.1038/s41467-023-37528-3

Figure Lengend Snippet: Fig. 2 | Interactions of TRIP4 variants in cells. a Western blots (WB) monitoring immunoprecipitation (IP) of ASCC1, ASCC2, and ASCC3 by the indicated N-terminally Flag-tagged TRIP4 variants from the cell extracts. b Western blots (WB) monitoring immunoprecipitation (IP) of ASCC3 by the indicated HA-tagged

Article Snippet: Western blotting was performed using antibodies against the Flag tag (Sigma-Aldrich F3165; 1:7500), ASCC1 (Proteintech 12301-1-AP; 1:500), ASCC2 (Proteintech 11529-1-AP; 1:1000) andASCC3 (Proteintech 17627- 1-AP; 1:1000).

Techniques: Western Blot, Immunoprecipitation

Journal: Nature communications

Article Title: Extended DNA threading through a dual-engine motor module of the activating signal co-integrator 1 complex.

doi: 10.1038/s41467-023-37528-3

Figure Lengend Snippet: Fig. 2 | Interactions of TRIP4 variants in cells. a Western blots (WB) monitoring immunoprecipitation (IP) of ASCC1, ASCC2, and ASCC3 by the indicated N-terminally Flag-tagged TRIP4 variants from the cell extracts. b Western blots (WB) monitoring immunoprecipitation (IP) of ASCC3 by the indicated HA-tagged

Article Snippet: Western blotting was performed using antibodies against the Flag tag (Sigma-Aldrich F3165; 1:7500), ASCC1 (Proteintech 12301-1-AP; 1:500), ASCC2 (Proteintech 11529-1-AP; 1:1000) andASCC3 (Proteintech 17627- 1-AP; 1:1000).

Techniques: Western Blot, Immunoprecipitation

Journal: bioRxiv

Article Title: Serine Racemase is a Cysteine Racemase and Physiologic Down Regulator of Insulin Promoter Methylation

doi: 10.1101/2022.05.17.492243

Figure Lengend Snippet: ( A ) HEK293 cells were transfected with ASCT2 and treated with different concentrations of D- cysteine (x axis) for 30 minutes at 37°C. Cells were harvested, washed and lysed and D-cysteine levels measured using luciferase assay. Data show levels of intracellular D- cysteine indicated by luminescence (y axis). ( B ) HEK293 cells were transfected with Asc-1 and treated with exogenous D-cysteine for 30 minutes at 37°C. Cells were harvested, washed and lysed and D-cysteine concentration measured using luciferase assay. Data show levels of intracellular D-cysteine indicated by luminescence. ( C ) Litter mate WT, Asc1 +/- and Asc1 -/- pups (10 day old) pancreas were harvested. Samples were pooled homogenized and sonicated to extract free endogenous D-cysteine and supernatants assayed using luciferase assay. Luminescence was measured in the supernatant samples and D-cysteine concentrations obtained from standard curve. Each genotype had N=10-12 mice. Data are representative of 3 independent experiments. ( D ) Pancreatic and liver lysates of WT and age-matched Asc1 -/- pups (litter mates) showing expression of SR. Actin was a loading control. (E) Schematic of D- cysteine transport in HEK293 cells by ASCT2 and Asc1 transporters.

Article Snippet: D-cysteine transport experiments were performed in HEK293 cells transiently transfected with human SLC1A15 (ASCT2; Origene; Cat# RC200305) and rat SLC7A10 (Asc1; Genomics-online.com; Cat# ABIN4047848) cDNA using lipofectamine 3000 reagent.

Techniques: Transfection, Luciferase, Concentration Assay, Sonication, Expressing

Journal: Frontiers in Oncology

Article Title: Inter and intra-tumor heterogeneity of paediatric type diffuse high-grade gliomas revealed by single-cell mass cytometry

doi: 10.3389/fonc.2022.1016343

Figure Lengend Snippet: Summary of the 15 antibodies used for the mass cytometry analysis.

Article Snippet: Anti-Human CD49c , 161Dy , Fluidigm , 3161016B , .

Techniques: Mass Cytometry