Journal: Cancer Science

Article Title: Antitumor effects of novel mAbs against cationic amino acid transporter 1 (CAT1) on human CRC with amplified CAT1 gene

doi: 10.1111/cas.14741

Figure Lengend Snippet: Correlation between gene amplification and mRNA expression of SLC7A1/CAT1 and CAT1 small interfering RNA (siRNA)–mediated growth inhibition of colorectal cancer (CRC) cells. A, Relative mRNA expression (ratio from a mean value of normal tissues) in normal and cancer tissues. B, Correlation between gene amplification and mRNA expression of SLC7A1. C, CAT1 mRNA levels in HCT116 and LS1034 cells treated with CAT1 siRNA for 24 hours. D, CRC cell growth 48 hours after treatment with CAT1 siRNA

Article Snippet: Human CRC cell lines (Caco2, COLO 201, COLO 205, COLO 320DM, HCT116, HT29, LS1034, LS123, LS174T, LS180, RKO, RKO‐E6, SNU‐C1, SW1116, WiDr), PK‐1, PK‐59, PANC‐1, MIA PaCa‐2 pancreatic, BT20, BT474 breast, A549, NCI‐H2170 lung cancers, and P3U1 mouse myeloma cells were purchased from the American Type Culture Collection (ATCC).

Techniques: Amplification, Expressing, Small Interfering RNA, Inhibition

Journal: Cancer Science

Article Title: Antitumor effects of novel mAbs against cationic amino acid transporter 1 (CAT1) on human CRC with amplified CAT1 gene

doi: 10.1111/cas.14741

Figure Lengend Snippet: Specificity of anti‐CAT1 monoclonal antibodies (mAbs) against human cancer cell lines. A, Representative flow cytometry (FCM) histograms of human colorectal cancer (CRC) cell lines stained with anti‐CAT1 mAbs. B and C, Different human CRC cell lines (B), and pancreatic, breast, and lung cancer cell lines (C) were reacted with CA2 or isotype‐matched control mAb, followed by incubation with phycoerythrin (PE)‐labeled anti‐rat IgG polyclonal antibody (pAb). The fluorescence intensity of individual cells was measured by FCM

Article Snippet: Human CRC cell lines (Caco2, COLO 201, COLO 205, COLO 320DM, HCT116, HT29, LS1034, LS123, LS174T, LS180, RKO, RKO‐E6, SNU‐C1, SW1116, WiDr), PK‐1, PK‐59, PANC‐1, MIA PaCa‐2 pancreatic, BT20, BT474 breast, A549, NCI‐H2170 lung cancers, and P3U1 mouse myeloma cells were purchased from the American Type Culture Collection (ATCC).

Techniques: Bioprocessing, Flow Cytometry, Staining, Control, Incubation, Labeling, Fluorescence

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Indirect effects of leptin receptor deficiency on lymphocyte populations and immune response in db/db mice.

doi: 10.4049/jimmunol.177.5.2899

Figure Lengend Snippet: FIGURE 5. Expression of OB-R in the thymus. A, Ob-Rb mRNA ex- pression was examined in the thymus 24 h after injection of saline (control) or 200 g/g hydrocortisone to WT C57BL/6 mice. Left panels, Represen- tative flow cytometric analyses of thymocytes from control (top) or hy- drocortisone (bottom)-injected mice stained for CD4 and CD8. Total thy- mocyte numbers were 45 106 cells (control) and 4 106cells (hydrocortisone). The right panels show Ob-rb (top) and actin (bottom) mRNA expression as assessed by RT-PCR. Lane 1, Control thymus; lane 2, hydrocortisone thymus; lane 3, negative control, PCR performed on non-reverse-transcribed RNA from control thymus. The position of mo- lecular mass markers (bp) is indicated on the right. B, Expression of WT and db alleles of Ob-rb mRNA was examined by RT-PCR on thymus RNA obtained from two db/db to WT BMCs. A schematic representation of the two alleles with the location of the PCR primers (arrows) is shown on the left. The db mutation () leads to insertion of an additional 106-bp exon (f), between exons 17 () and 18b (u) of the mouse lepr gene. This additional exon contains a stop codon causing premature termination of the Ob-rb protein in the db mutant (42). Amplification of the WT and db alleles with the indicated primers yields PCR products of 170 and 276 bp, respec- tively. The right panel shows the expression of mRNA encoding both WT (recipient-derived) and db (donor-derived) alleles in the thymus of two different db/db to WT BMCs (lanes 1 and 2). C, Expression of the Ob-rb protein was examined by Western blotting in thymocytes, whole thymus, a thymic stromal fraction, and in the thymic epithelial cell line MTE-4.14 (upper panel). The membrane was stripped and reblotted with anti--actin Abs (lower panel). The positions of molecular mass standards are indicated on the right. Lane 1, Human COLO 320DM cell lysate (positive control); lane 2, total thymocytes; lane 3, whole thymus; lane 4, thymic stromal fraction; and lane 5, MTE- 4.14 total cell lysate.

Article Snippet: Human COLO 320DM whole cell lysate (Santa Cruz Biotechnology) was used as a positive control for OB-R expression.

Techniques: Expressing, Injection, Saline, Control, Staining, Reverse Transcription Polymerase Chain Reaction, Negative Control, Reverse Transcription, Mutagenesis, Derivative Assay, Western Blot, Membrane, Positive Control