Science - USA (2021-07-16)

XhoI and EcoRI. The entire mnGFP-minD fusion protein was cloned into thePBAD-GFP po- sition (XbaI and SphiI) of pFcCGi, creating ...

withStmwere performed on individual tripli- cates at MOI 5. After 5 hours, cells were washed three times in sterile prewarmed DM ...

dialyzed extensively (10 kDa MWCO) into 20 mM acetic acid to both refold the protein and remove unincorporated dye. Guanylate bi ...

and incubated for 10 min. Bacteria were washed 3× with PBS containing 0.2% glucose and 1% dimethylformamide (DMF). 500 nM rAPOL3 ...

glow-discharged (PELCO easiGlow Glow Dis- charge Cleaning System) for 20 s to increase hydrophilicity of the carbon surface. 5-m ...

L. Vanhammeet al., Apolipoprotein L-I is the trypanosome lytic factor of human serum.Nature 422 , 83–87 (2003). doi:10.1038/nat ...

RESEARCH ARTICLE SUMMARY ◥ IMMUNOLOGY Expression of Foxp3 by T follicular helper cells in end-stage germinal centers Johanne T. ...

RESEARCH ARTICLE ◥ IMMUNOLOGY Expression of Foxp3 by T follicular helper cells in end-stage germinal centers Johanne T. Jacobsen ...

individual GCs as they approached their dis- solution phase (Fig. 2, A and B; fig. S2A; and movie S1). Longitudinal imaging show ...

and are detectable using our intravital imaging setup. Imaging of early (day 10) prime-boost GCs generated as in Fig. 1A reveale ...

the same experiment longitudinally using an iLN imaging window. Corroborating our previous data, Foxp3 expression began to rise ...

“mixed”clones remained predominantly with- in TFHclusters 1 and 2 and were not found within cluster 4, likely to include most of ...

expression of Foxp3 in peak GC TFHcells led to a roughly 60% reduction in GC size com- pared with that in control mice that did ...

least has the potential to play a role inFoxp3 up-regulation by TFHcells. Another potential factor is the availability of antige ...

Jacobsenet al.,Science 373 , eabe5146 (2021) 16 July 2021 8 of 13 F Foxp3+ vs. Foxp3– cells in mixed clones A Photoactivated T c ...

to Alexa Fluor 633 or Alexa Fluor 594 (Thermo Fisher), 24 hours before imaging ( 52 ). For photoactivation experiments, FDCs wer ...

were placed under the microscope objective, connected to an objective heater set to 40°C. Longitudinal iLN window imaging was pe ...

T cell culture Foxp3-RFP–TFHand non-TFHCD4+T cells were sorted from Foxp3IRES-RFPreporter mice immunized with NP-OVA in alhydrog ...

S. Crotty, Follicular helper CD4 T cells (TFH).Annu. Rev. Immunol. 29 , 621–663 (2011). doi:10.1146/annurev-immunol- 031210-101 ...

input from S.J.A. and A.K.S.Competing interests:A.K.S. reports compensation for consulting and/or scientific advisory board (SAB ...