Journal: bioRxiv

Article Title: YAP1 status defines two intrinsic subtypes of LCNEC with distinct molecular features and therapeutic vulnerabilities

doi: 10.1101/2023.12.19.572449

Figure Lengend Snippet: YAP1 status does not predict sensitivity to cisplatin in LCNEC cell lines (A) or cell line/PDX xenograft models (B). C, Treatment of LCNEC cell lines with DMSO, cisplatin, MYF-01-37, XAV-939 and decitabine did not change YAP1, but verteporfin reduced YAP1 levels. D, YAP1 status does not predict response to verteporfin. E, Waterfall plot of drug sensitivity in LCNEC cell lines. A comparison of high YAP1 levels and IC50 values identified several drugs with similar targets, including MEK1/2, CDK4/6, and Src family kinase inhibitors. F, Treatment of YAP1-high LCNEC and SCLC (SW1271) cell lines with trametinib did not change YAP1 levels. G, Tumor growth curves from YAP1-high cell line xenografts H1915 (LCNEC) and SW1271 (SCLC) demonstrate a delay in tumor growth with trametinib treatment.

Article Snippet: One million cells each for H810, H1155, H1755, H1833, H2106, MKL1, HCC2374, HCC4017, HOP92, H1359, H2066, H1770, H2106, H1570, H661, HCC3051, H460, HCC4017, H1299, and H1915 in triplicate were surfaced stained with DLL3 (FAB4315P; R&D Systems), AXL (386202; Biolegends), CD56 (362534), EPCAM (324222) or IgG control and then fixed in 2% PFA.

Techniques: Comparison

Journal: The Journal of biological chemistry

Article Title: The plastid-encoded ccsA gene is required for heme attachment to chloroplast c-type cytochromes.

doi: 10.1074/jbc.271.9.4632

Figure Lengend Snippet: FIG. 5. Phenotype of a strain (DccsA) containing a disrupted ccsA gene. Soluble proteins were prepared from copper-supplemented (1Cu) or copper-deficient (2Cu) cultures of the indicated strains and analyzed for the accumulation of holocytochrome c6 (heme stain and anti-cytochrome c6) and plastocyanin (anti-pc). Nitrocellulose membranes were used for the leftmost three panels. Bound antibody was detected by use of a horseradish peroxidase-conjugated secondary antibody. The pellet fractions from the preparation were analyzed for cytochrome f accumulation (anti-cytochrome f). PVDF membranes were used for the transfer, and an alkaline phosphatase-conjugated secondary antibody was used for detection.

Article Snippet: Bound antibodies were detected with alkaline phosphatase- or horseradish peroxidase-conjugated secondary antibody according to instructions provided by the manufacturer (Bio-Rad).

Techniques: Staining

Journal: The Journal of biological chemistry

Article Title: The plastid-encoded ccsA gene is required for heme attachment to chloroplast c-type cytochromes.

doi: 10.1074/jbc.271.9.4632

Figure Lengend Snippet: FIG. 6. Complementation of strain B6 with the cloned ycf5 gene. A, sum- mary of the complementation experi- ments. Plasmids pEBP and pEBH were constructed in vector pTZ19R, while pNH was constructed in the vector pET22b(1). The indicated frequencies are the aver- ages from three different experiments. B, extracts of soluble protein were prepared from copper-deficient cultures of strain CC425 (wt), strain B6, or cells of B6 res- cued by plasmid pEBP (B6-P) or pEBH (B6-H). Equivalent amounts (correspond- ing to 1 A595 unit in the Pierce Coomassie dye binding assay) were analyzed, after separation of proteins in an SDS-contain- ing polyacrylamide gel and transfer to a nitrocellulose membrane, for accumula- tion of holocytochrome c6 by heme stain- ing (bottom panel, 45-min exposure to NEN Reflection film) or decoration with anti-cytochrome c6 (top panel, horse- radish peroxidase conjugated second antibody).

Article Snippet: Bound antibodies were detected with alkaline phosphatase- or horseradish peroxidase-conjugated secondary antibody according to instructions provided by the manufacturer (Bio-Rad).

Techniques: Clone Assay, Construct, Plasmid Preparation, Binding Assay, Membrane, Staining

Journal: The Journal of biological chemistry

Article Title: Identification of quinone-binding and heme-ligating residues of the smallest membrane-anchoring subunit (QPs3) of bovine heart mitochondrial succinate:ubiquinone reductase.

doi: 10.1074/jbc.274.13.8717

Figure Lengend Snippet: FIG. 1. Identification of recombinant QPs3 by Western blot. A, SDS-PAGE of succinate:ubiquinone reductase (lane 2), recombinant GST-QPs3 fusion protein (lane 3), thrombin-treated fusion protein (lane 4), purified recombinant QPs3 (lane 5). The molecular mass standard containing phosphorylase B (108 kDa), bovine serum albumin (84 kDa), ovalbumin (53 kDa), carbonic anhydrase (35 kDa), soybean trypsin inhibitor (28 kDa), and lysozyme (20 kDa) is in the lane 1. The proteins in A were electrophoretically transferred to nitrocellulose membrane and reacted with anti-QPs3 peptide antibodies (B) and anti-GST-QPs3 antibodies (C). Protein A horseradish peroxidase conjugate was used as a second antibody.

Article Snippet: Agarose, acrylamide, bisacrylamide, horseradish peroxidase color development reagent, protein A horseradish peroxidase conjugate, and Bradford reagent were obtained from Bio-Rad.

Techniques: Recombinant, Western Blot, SDS Page, Purification, Membrane