Journal: bioRxiv

Article Title: Kinetic and functional analysis of abundant microRNAs in extracellular vesicles from normal and stressed cultures of Chinese Hamster Ovary (CHO) cells

doi: 10.1101/2023.06.21.545902

Figure Lengend Snippet: (a) Cell growth curve of cells transiently expressing let-7a, miR-21, or miR-92a compared to the control (transient transfection of pLifeAct-miRFP703). (b) Annexin V/7AAD flow cytometry viability assay of cultures transiently transfected with the control plasmid (pLifeAct-miRFP703), let-7a plasmid, miR-21 plasmid, or miR-92a plasmid. Conditions where viability was significantly lower than the associated control are red. Conditions where viability was significantly greater than the associated control are green. (c) Fold change in cultures transiently expressing let-7a, miR-21, and miR-92a compared to the endogenous expression in the control culture (transient transfection of pLifeAct-miRFP703). Error bars represent the standard error of the mean (SEM) of 3-4 replicates. Significance relative to control was determined with unpaired Student’s t-test, * for p<0.05, ** for p<0.01, *** for p<0.001.

Article Snippet: CHO cells were electroporated with 2 μg of plasmids containing the primary or a fragment of the primary microRNA sequences of three miRs that were in high abundance in standard or stressed cultures (miR-21 (Addgene #21114), miR-92a (Addgene #46672), let-7a (Addgene #51377) using the Nucleofector V kit (Lonza, Basel, Switzerland) according to manufacturer’s protocols.

Techniques: Expressing, Transfection, Flow Cytometry, Viability Assay, Plasmid Preparation

Journal: bioRxiv

Article Title: Kinetic and functional analysis of abundant microRNAs in extracellular vesicles from normal and stressed cultures of Chinese Hamster Ovary (CHO) cells

doi: 10.1101/2023.06.21.545902

Figure Lengend Snippet: (a) Cell growth curve of cells transiently expressing let-7a, miR-21, or miR-92a compared to the control (transient transfection of pLifeAct-miRFP703). (b) Annexin V/7AAD flow cytometry viability assay of cultures transiently transfected with the control plasmid (pLifeAct-miRFP703), let-7a plasmid, miR-21 plasmid, or miR-92a plasmid. Conditions where viability was significantly lower than the associated control are red. Conditions where viability was significantly greater than the associated control are green. (c) Fold change in cultures transiently expressing let-7a, miR-21, and miR-92a compared to the endogenous expression in the control culture (transient transfection of pLifeAct-miRFP703). Error bars represent the standard error of the mean (SEM) of 3-4 replicates. Significance relative to control was determined with unpaired Student’s t-test, * for p<0.05, ** for p<0.01, *** for p<0.001.

Article Snippet: CHO cells were electroporated with 2 μg of plasmids containing the primary or a fragment of the primary microRNA sequences of three miRs that were in high abundance in standard or stressed cultures (miR-21 (Addgene #21114), miR-92a (Addgene #46672), let-7a (Addgene #51377) using the Nucleofector V kit (Lonza, Basel, Switzerland) according to manufacturer’s protocols.

Techniques: Expressing, Transfection, Flow Cytometry, Viability Assay, Plasmid Preparation