fetal bovine serum fbs  (Thermo Fisher)

Journal: PLoS Pathogens

Article Title: Establishment of Human Papillomavirus Infection Requires Cell Cycle Progression

doi: 10.1371/journal.ppat.1000318

Figure Lengend Snippet: Cell cycle arrest by serum starvation abrogates HPV infection. (A) After 24 hr incubation in serum-free DMEM, HaCaT cells were inoculated with HPV pseudovirion hpv 16wpA-RL, and RL activity was measured after 2 d. For serum conversion, serum-free DMEM was replaced with 10% FBS/DMEM before virus inoculation. The expression value is shown as % infectivity to untreated cells with hpv 16wpA-RL inoculation. Columns , mean; bars , SD. (B) Flow cytometry was performed to confirm cell cycle arrest by serum starvation. The solid line indicates the HaCaT cells incubated with serum-free DMEM for 24 h, and the gray shaded area indicates control HaCaT cells in DMEM with 10% FBS.

Article Snippet: Cell lines Human embryonic kidney cell line 293T from ATCC, and its enhanced SV40 T antigen-expressing daughter cell line 293TT from John Schiller, were maintained in Dulbecco's modified eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Invitrogen).

Techniques: Infection, Incubation, Activity Assay, Expressing, Flow Cytometry, Cytometry

Journal: PLoS Pathogens

Article Title: Establishment of Human Papillomavirus Infection Requires Cell Cycle Progression

doi: 10.1371/journal.ppat.1000318

Figure Lengend Snippet: Cell cycle arrest does not inhibit influenza virus. (A) After 24 hr synchronization with aphidicolin and 4 hr treatment with etoposide or aphidicolin (3 µM each), 293T cells were inoculated in parallel with hpv 16wpA-RL or an influenza virus vector in which the hemagglutinin and neuraminidase open reading frames in viral RNA were replaced with those of vesicular stomatitis virus glycoprotein and RL, respectively [50] . (B) After 24 hr synchronization in serum-free DMEM, HaCaT cells were inoculated with hpv 16wpA-RL and the RL-expressing influenza virus as (A) in the presence or absence of FBS. RL activity was measured after 48 hrs, as described in Materials and Methods , normalized to RL activity in equivalently inoculated cells maintained in 10% FBS, and expressed in the histograms as % infectivity. Columns , mean; bars , SD.

Article Snippet: Cell lines Human embryonic kidney cell line 293T from ATCC, and its enhanced SV40 T antigen-expressing daughter cell line 293TT from John Schiller, were maintained in Dulbecco's modified eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Invitrogen).

Techniques: Plasmid Preparation, Expressing, Activity Assay, Infection

Journal: Stem Cell Reviews

Article Title: Improved Approach for Chondrogenic Differentiation of Human Induced Pluripotent Stem Cells

doi: 10.1007/s12015-014-9581-5

Figure Lengend Snippet: Chondrogenic differentiation of hiPSC. ( a ) Classical chondrogenic differentiation of hiPSCs via formation of embryoid bodies, outgrowth of endodermal ( green ), ectodermal ( yellow ) and mesodermal ( red ) cell lineages, selection of mesodermal cells, and induction of MSC and induction of chondrocytes. In this method hiPS cells were detached from matrigel coated dish and moved to ultra low attachment culture dish for 5 days to induce the EB formation, then EBs moved to plastic culture dish to select the hMSCs by collecting the outgrowing cells from EB (from day 5 to day 14) after collecting the attached fibroblast-like cells. These cells were cultured for 3 weeks in media containing FBS to prepare the hiPSC-MSCs (day 35 of differentiation). Then, hiPSC-MSCs were differentiated in a pellet culture system using serum free chondrogenic media for 3 weeks. ( b ) Embryoid body free method of direct differentiation of hiPSCs into hiPSC-MSCs, followed by chondrogenic differentiation. In embryoid body free method hiPSCs were cultured in matrigel coated dish and media was changed to hMSC media (DMEM supplemented with FBS) for 5 days to induce the hMSC differentiation (Day 5). Then, cells were detached and moved to a plastic culture dish for 4 passages to prepare the hiPSC-MSCs (Day 28). To differentiate the hiPSC-MSCs to chondrocytes, cells were used in pellet culture system using serum free chondrogenic media for 3 weeks

Article Snippet: Osteogenic differentiation was induced by culturing 6 × 104 cells/cm2 in osteogenic differentiation medium consisting of DMEM supplemented by 10 % FBS (Gibco), 100 U/ml penicillin, 100 μg/ml streptomycin (Gibco), 10 % L-Glutamine (Gibco), 50 μg/ml L-ascorbic acid 2-phosphate sequimagnesium (Sigma), 100 μg/ml MEM sodium pyruvate (Gibco), 0.1 μM dexamethasone (Sigma), and 100 mM b-glycerophosphate.

Techniques: Selection, Cell Culture

Journal: PLoS ONE

Article Title: Molecular Characterization and Transcriptional Regulation Analysis of the Bovine PDHB Gene

doi: 10.1371/journal.pone.0157445

Figure Lengend Snippet: Promoter activity analysis of the bovine PDHB gene. a. We transferred six serial deletion constructs in pGL3-basic into C2C12 cells. After 5 h we replaced the transfection mixture with DMEM with 5% FBS (myoblasts) or 2% HS (myotubes). b. We transferred the same constructs into 3T3-L1 cells. We normalized relative luciferase activities to Renilla luciferase activity. The transcription factor binding sites of MYOG and C/EBPß are indicated with closed circles and ellipses, respectively. *, P

Article Snippet: Cell culture, transfection and dual-luciferase reporter assay We cultured the mouse myoblast cell line (C2C12) and the 3T3-L1 cell line in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (GIBCO-Invitrogen) under humidified air containing 5% CO2 at 37°C.

Techniques: Activity Assay, Construct, Transfection, Luciferase, Binding Assay

Journal: Molecular Biotechnology

Article Title: Ribosome Display Selection of a Murine IgG1 Fab Binding Affibody Molecule Allowing Species Selective Recovery Of Monoclonal Antibodies

doi: 10.1007/s12033-010-9367-1

Figure Lengend Snippet: Binding studies of Z mab25 or protein G with fetal bovine serum and species selective affinity recovery of two monoclonal antibodies. a 10% FBS was flowed over a Z mab25 ( gray solid trace ) or protein G C2–C3 fragment (SPG) ( black dashed trace ) BIAcore biosensor surface. For comparison, a sample containing 10% FBS spiked with mouse IgG 1 mAb3 was also flowed over the Z mab25 surface ( black solid trace ). b : SDS-PAGE analysis (reducing conditions) of samples, flow-through (FT), and eluate (E) fractions from affinity chromatography experiments using either a protein G (SPG) column or a Z mab25 column, and samples of 10% FBS only ( lanes 1 – 6 ) or 10% FBS with in-spiked mouse IgG 1 mAb3 ( lanes 7 – 12 ) or 10% FBS with in-spiked mouse IgG 1 8F11 ( lanes 13 – 15 ). M LMW-SDS molecular weight marker; lane 1 : 10% FBS sample; lane 2 : SPG column, FT; lane 3 : SPG column, E; lane 4 : 10% FBS sample; lane 5 : Z mab25 column, FT; lane 6 : Z mab25 column, E; lane 7 : 10% FBS + mAb3 sample; lane 8 : SPG column, FT; lane 9 : SPG column, E; lane 10 : 10% FBS + mAb3 sample; lane 11 : Z mab25 column, FT; lane 12 : Z mab25 column, E; lane 13 : 10% FBS + mAb 8F11 sample; lane 14 : Z mab25 column, FT; lane 15 : Z mab25 column, E. The black arrows to the right indicate nominal molecular weights of antibody heavy and light chains, respectively. The numbers to the left indicate molecular weights in kilo Dalton (kDa). c Results from a biosensor analysis of pH neutralized eluates from the SPG and Z mab25 columns, originating from the use of the 10% FBS sample containing in-spiked mAb3 ( 1 – 4 ) or mAb 8F11 ( 5 , 6 ). On separate flow cell surfaces, anti-cow IgG and anti-mouse Ig antibodies were immobilized, and samples were injected. (1) black solid trace SPG column eluate, anti-cow IgG surface; (2) black dashed trace SPG column eluate, anti-mouse Ig surface; (3) gray solid trace Z mab25 column eluate, anti-cow IgG surface; (4) gray dashed trace Z mab25 column eluate, anti-mouse Ig surface; (5) black dotted trace Z mab25 column eluate, anti-cow IgG surface; (6) gray dotted trace Z mab25 column eluate, anti-mouse Ig surface

Article Snippet: For analysis of Zmab25 or protein G C2–C3 domain interaction with fetal bovine serum (FBS), a third CM5 sensor chip was prepared onto which 488 RU of protein G C2–C3 and 1817 RU of Zmab25 were immobilized on separate surfaces, and 100 μl of 10% FBS (Gibco) or 10% FBS and 133 nM mAb3 were flowed over both surfaces.

Techniques: Binding Assay, SDS Page, Flow Cytometry, Affinity Chromatography, Molecular Weight, Marker, Injection