Journal: Developmental Cell

Article Title: The Ciliary Machinery Is Repurposed for T Cell Immune Synapse Trafficking of LCK

doi: 10.1016/j.devcel.2018.08.012

Figure Lengend Snippet: ARL3-Specific Release Localizes LCK to the Immune Synapse (A and F) T:B cell conjugates stained with anti ARL3, LCK, or ARL13B antibodies as indicated in the presence or absence of SEE. (B) LR of ARL3, ARL13B, and LCK of conjugates from (A) and (F): ARL3 shows no localization to the synapse in the presence (LR = 0.3) or absence of SEE (n = 38). LCK shows LR = 1.9 in the presence of SEE and 0.7 in its absence (n = 42; p ≤ 0.0001). ARL13B shows LR = 2 in the presence of SEE (n = 32) and 0.6 in its absence (n = 16; p ≤ 0.0001). (C) ARL3 WT GFP and the constitutively active ARL3 Q71L GFP were nucleofected into cells and T:B cell conjugates, in the presence of SEE, were stained with anti LCK antibodies. (D) LR of LCK of conjugates from (C). ARL3 WT GFP expression had little effect on LR of LCK (n = 18) compared to cells not expressing ARL3 WT GFP (LR = 2). ARL3 Q71L GFP-expressing cells showed LR of LCK = 1 (n = 16, p = 0.0002). Differences in LR were analyzed using the Mann-Whitney U test (B and D). (E) CD69 expression was analyzed via flow cytometry for cells stimulated with anti CD28 and CD3 antibodies and overexpressing ARL2-GFP, ARL2 Q70L GFP, ARL3 Q71L GFP, ARL3-GFP, or GFP. ARL3 Q71L GFP (p ≤ 0.0001), ARL3 WT GFP (p = 0.004), ARL2 WT GFP (p = 0.02), and ARL2 Q70L GFP (p = 0.03) p values refer to unpaired t test data. (G) Ciliated RPE cell stained with anti ARL13B antibody. Scale bars, 5 μm.

Article Snippet: Cells were stimulated for 4 hours and then analysed using flow cytometry to identify CD69 expression (CD69APC, Invitrogen, USA, MHCD6905).

Techniques: Staining, Expressing, MANN-WHITNEY, Flow Cytometry, Cytometry

Journal:

Article Title: The symbiosis regulator RscS controls the syp gene locus, biofilm formation and symbiotic aggregation by Vibrio fischeri

doi: 10.1111/j.1365-2958.2006.05475.x

Figure Lengend Snippet: Transcription of syp genes by rscS . After 24 h growth at 22°C in HMM, β-galactosidase activity levels were determined for syp :: lacZ reporter strains carrying

Article Snippet: Transconjugants were selected for the presence of the plasmid and screened for induction of β-galactosidase expression on SWT Tet containing 80 μg ml−1 X-gal (Molecular Probes).

Techniques: Activity Assay

Journal: Immunology

Article Title: Cord blood mesenchymal stem cells propel human dendritic cells to an intermediate maturation state and boost interleukin-12 production by mature dendritic cells

doi: 10.1111/j.1365-2567.2009.03142.x

Figure Lengend Snippet: CCR7 expression is up-regulated in mature dendritic cells (mDCs) co-cultured with unrestricted somatic stem cells (USSCs). DCs obtained from monocytes after 6 days of induction with granulocyte–macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) [immature DCs (iDCs)] were stimulated by lipopolysaccharide (LPS) for an additional 48 hr (mDC) with or without USSC co-culture. CCR7 expression levels were determined by flow cytometry. The results are percentage of positive cells (± standard error of the mean) of triplicate data ( n = 3).

Article Snippet: In the migration studies, CCR7 expression (anti-human CCR7, MAB197; R & D Systems, Minneapolis, MN) was analyzed in the same manner as described above.

Techniques: Expressing, Cell Culture, Co-Culture Assay, Flow Cytometry, Cytometry

Journal: Molecular Oncology

Article Title: A novel enediyne‐integrated antibody–drug conjugate shows promising antitumor efficacy against CD30+ lymphomas

doi: 10.1002/1878-0261.12166

Figure Lengend Snippet: Receptor‐mediated internalization of anti‐CD30‐LDP by CD30 + lymphoma cells. Cells were treated at 4 °C with anti‐CD30‐LDP labeled with Alexa Fluor 488‐conjugated anti‐human IgG. Anti‐CD30‐LDP bound to CD30‐positive cell line L540 and Karpas299, while no signal was detected on CD30‐negative cell line Raji (A). Anti‐CD30‐LDP was internalized and trafficked to the lysosomes in L540 (B) and Karpas299 (C) cells at 37 °C for 24 h. Cell surface or intracellular anti‐CD30‐LDP was visualized by immunofluorescence confocal microscopy. Anti‐CD30‐LDP is shown in green, the lysosomal marker Lamp‐1 is shown in red, and the Hoechst‐stained nuclei are in blue. Colocalization of signals for anti‐CD30‐LDP with Lamp‐1 is shown in yellow. The data shown were representative of three independent experiments. Scale bar = 10 μm.

Article Snippet: For the generation of anti‐CD30‐LDP and anti‐CD30 antibody‐expressing cell lines, pIZDHL‐anti‐CD30‐LDP and pIZDHL‐anti‐CD30 were linearized and transfected into CHO/dhFr‐ cells by lipofectin transfection (Invitrogen, Carlsbad, CA, USA), respectively.

Techniques: Labeling, Immunofluorescence, Confocal Microscopy, Marker, Staining

Journal: Molecular Oncology

Article Title: A novel enediyne‐integrated antibody–drug conjugate shows promising antitumor efficacy against CD30+ lymphomas

doi: 10.1002/1878-0261.12166

Figure Lengend Snippet: In vivo efficacy of anti‐CD30‐LDM against Karpas299 xenograft model. (A) The antitumor effects of anti‐CD30‐LDM in NOD/SCID mice bearing Karpas299 xenografts ( n = 6). The agents were administered at doses indicated in the figure on a Q7D × 2 schedule, and arrows indicate days of administration. * P

Article Snippet: For the generation of anti‐CD30‐LDP and anti‐CD30 antibody‐expressing cell lines, pIZDHL‐anti‐CD30‐LDP and pIZDHL‐anti‐CD30 were linearized and transfected into CHO/dhFr‐ cells by lipofectin transfection (Invitrogen, Carlsbad, CA, USA), respectively.

Techniques: In Vivo, Mouse Assay

Journal: Molecular Oncology

Article Title: A novel enediyne‐integrated antibody–drug conjugate shows promising antitumor efficacy against CD30+ lymphomas

doi: 10.1002/1878-0261.12166

Figure Lengend Snippet: Targeting of fluorescently labeled proteins to established L540 and Karpas299 xenografts in NOD/SCID mice. Anti‐CD30‐LDP (A) or free LDP (B) was labeled with DyLight 680 and injected into tumor‐bearing NOD/SCID mice through tail veins at 20 mg·kg −1 , respectively. In vivo fluorescence images were taken at appointed time. Color scale represents photons·s −1 ·cm −2 ·steradian −1 .

Article Snippet: For the generation of anti‐CD30‐LDP and anti‐CD30 antibody‐expressing cell lines, pIZDHL‐anti‐CD30‐LDP and pIZDHL‐anti‐CD30 were linearized and transfected into CHO/dhFr‐ cells by lipofectin transfection (Invitrogen, Carlsbad, CA, USA), respectively.

Techniques: Labeling, Mouse Assay, Injection, In Vivo, Fluorescence