Journal: Cancer Immunology Research

Article Title: MicroRNAs Affect Complement Regulator Expression and Mitochondrial Activity to Modulate Cell Resistance to Complement-Dependent Cytotoxicity

doi: 10.1158/2326-6066.cir-18-0818

Figure Lengend Snippet: Figure 6. Modulation of the membrane complement regulatory proteins by microRNAs. The expression of CD46 (A), CD55 (B), and CD59 (C) in lysates of K562 cells transfected with microRNA inhibitors or with control, determined by the proteomic analysis described in Materials and Methods, is shown as the mean SD of log2 of intensity value units of five independent experiments. , P < 0.05; , P < 0.01, relative to control (Student t test). NS, not significant. D, K562 cells were transfected with miR-616 inhibitor plasmid or with control (C) plasmid as a negative control. After 48 hours, the cells were labeled with anti-CD46, anti-CD55, or anti-CD59 and then with fluorescently labeled secondary antibody. Cells were then analyzed by flow cytometry, and the mean fluorescence intensity (MFI) values, representative of three independent experiments, were determined. The expression of each regulator in anti–miR-616–treated cells was normalized to their levels in control cells (set as 100). , P < 0.05; , P < 0.01, relative to control (Student t test). E–G, K562 cells were transfected with a miR-150 expression plasmid or a control plasmid. After 24 hours, the cells were labeled with mouse anti-CD46 (E), anti-CD55 (F), or anti-CD59 (G) and fluorescently labeled secondary antibody. Cells were then analyzed by flow cytometry, and MFI values, representative of three independent experiments, were determined. , P < 0.05, relative to control (Student t test).

Article Snippet: Mouse monoclonal antihuman CD46 (clone MEM-258), anti-human CD55 (clone 67), and anti-human CD59 (clone MEM-43) antibodies were purchased from AbD Serotec.

Techniques: Membrane, Expressing, Transfection, Control, Plasmid Preparation, Negative Control, Labeling, Cytometry

Journal:

Article Title: Distinct Proinflammatory Host Responses to Neisseria gonorrhoeae Infection in Immortalized Human Cervical and Vaginal Epithelial Cells

doi: 10.1128/IAI.69.9.5840-5848.2001

Figure Lengend Snippet: Immunocytochemical analysis of adhesion molecule expression by cervicovaginal epithelial cells in vitro and in vivo. Positive cells appear red. (A and B) Constitutive expression of CD46 in endocervical tissue (A) and endocervical cell culture (B). (C to G) Expression of CD66 in vaginal tissue (C), ectocervical tissue (D), endocervical tissue (E), uninfected endocervical cell culture (F), and endocervical cell culture after 8 h of infection with the N. gonorrhoeae F62 piliated variant (G). (H to K) Expression of ICAM-1 in endocervical cell cultures with no infection (H), following 8 h of TNF-α stimulation (I), following 8 h of infection with N. gonorrhoeae piliated F62 (J), or following 8 h of infection with N. gonorrhoeae nonpiliated F62 (K). L. lumenal epithelial surface: B. basal epithelial layers. Magnification, ×125 (A, C to E, and H to K) and ×250 (B, F, and G).

Article Snippet: Anti-human CD46 antibody (mouse monoclonal MAS657) was purchased from Accurate Chemical and Scientific Corporation, Westbury, N.Y. Fluorescein isothiocyanate (FITC)-conjugated pan-CD66 antibody (mouse monoclonal F7112) recognizing an epitope common to four members of the CD66 family (CD66a, -b, -c, and -e) was purchased from Dako, Carpinteria, Calif.

Techniques: Expressing, In Vitro, In Vivo, Cell Culture, Infection, Variant Assay