Lentivirus-mediated B cell transduction

Lentivirus preparation and B cell transduction were described previously.^{59 (link)} Briefly, 293T cells were cotransfected with psPax2 plasmid (Addgene), VSVG plasmid (Addgene), siControl plasmid, siHSP70 plasmid or siHSF1 plasmid (ABM, Richmond, BC, Canada), lenti-EV (empty vector) or lenti-HSP70 overexpression plasmid (Sinobiological) using Lipofectamine 2000 (Thermo Fisher). Supernatants were collected and filtered. Isolated bone marrow B cells were transduced by lentiviral particles by spin infection.

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Meng X., Lin Z., Cao S., Janowska I., Sonomoto K., Andreev D., Katharina K., Wen J., Knaup K.X., Wiesener M.S., Krönke G., Rizzi M., Schett G, & Bozec A. (2022). Estrogen-mediated downregulation of HIF-1α signaling in B lymphocytes influences postmenopausal bone loss. Bone Research, 10, 15.

Publication 2022

psPax2 plasmid VSVG plasmid Lipofectamine 2000

293t cells B cells Bone marrow Hsp70 Infection Lentivirus Lipofectamine 2000 Plasmid Vector

Corresponding Organization :

Other organizations : Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, University Medical Center Freiburg, University of Freiburg

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Variable analysis

independent variables

SiControl plasmid
SiHSP70 plasmid
SiHSF1 plasmid
Lenti-EV (empty vector)
Lenti-HSP70 overexpression plasmid

dependent variables

Not explicitly mentioned

control variables

PsPax2 plasmid (Addgene)
VSVG plasmid (Addgene)
Lipofectamine 2000 (Thermo Fisher)

controls

Negative control: siControl plasmid
Positive control: lenti-HSP70 overexpression plasmid

Annotations

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