Journal: PLoS ONE

Article Title: Application of JC1 for non-toxic isolation of cells with MDR transporter activity by flow cytometry

doi: 10.1371/journal.pone.0174905

Figure Lengend Snippet: All cells were incubated for 60 or 75 min with either 250 nM JC1 or 4 μg/ml Hoechst, unless indicated otherwise. Inhibitors were added 60 min before the addition of JC1 or Hoechst. The % of cells included within subjectively selected JC1-SP or Hoechst-SP domains (gates) are indicated next to the gate. A and B. Sixty min incubation with JC1; control (Cntrl) and puromycin-selected cells transduced with the ABCG2-A shRNA. C-E. Seventy five min incubation with JC1 incubated for 75 min; Control (C) and ABCG2 gene transduced cells before (D) or after (E) antibiotic selection. Note that the shRNA, in addition to the elimination of the JC1 low cohort, causes a rightward shift of JC1 main cells (B) while conversely, addition of the ABCG2 gene causes not only a large increase in the JC1 low but also and a leftward shift of JC1 main cells (E). F-H. Hoechst stain for 75 min. F. C ontrol cells. G. Cells pre-incubated with 2 μM Ko143. The G0/G1, S and G2-M cell cohorts are indicated. H. ABCG2 shRNA transduced and puromycin selected cells. I. Western blot of unmodified HCEC and HCEC transduced with ABCG2 shRNA (shRNA) or ABCG2 gene. Top: ABCG2 stain. Bottom: GAPDH stain of the same blot.

Article Snippet: A lentiviral plasmid for the expression of ABCG2 (pSIN4-EF2-ABCG2-IRES-Neo) was obtained from Addgene.

Techniques: Incubation, Control, Transduction, shRNA, Selection, Staining, Western Blot

Journal: PLoS ONE

Article Title: Application of JC1 for non-toxic isolation of cells with MDR transporter activity by flow cytometry

doi: 10.1371/journal.pone.0174905

Figure Lengend Snippet: Cells were incubated with dyes for 75 min. A-C. Control (A), and ABCB1shRNA transduced (B) or and scrambled shRNA transduced cells (C). D-F . Control cells and cells transduced with the ABCG2-A or ABCG2-B shRNAs, respectively. G-I. Control medium (G), medium complemented with 2 μM of either valspodar (H) or Ko143 (I), respectively. J and K. Hoechst 33342 stain in control (J) or after transduction with the ABCG2-A gene (K), respectively. L. Flow cytometry histogram of control or ABCB1shRNA-transduced live 293T immunostained with FITC-conjugated anti-ABCB1 antibody. An FITC conjugated isotype control was used to confirm the specificity of the staining.

Article Snippet: A lentiviral plasmid for the expression of ABCG2 (pSIN4-EF2-ABCG2-IRES-Neo) was obtained from Addgene.

Techniques: Incubation, Control, shRNA, Transduction, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: Application of JC1 for non-toxic isolation of cells with MDR transporter activity by flow cytometry

doi: 10.1371/journal.pone.0174905

Figure Lengend Snippet: A-C. Control (A), ABCB1-A shRNA (B) and ABCG2-A shRNA transduced (C) cells were incubated with JC1 for 75 min. D-I. Control cells (D) and cells pretreated with 2 μM Ko143 (E), 5 μM Ko143 (F), 2 μM FTC (G), 2 μM Valspodar (H), or 5 μM valspodar (I) were incubated with JC1 for 90 min. J-L. Control cells (J), and cells pretreated with 2 μM Ko143 (K) or transduced with ABCB1-A shRNA (L) were incubated with Hoechst for 75 min.

Article Snippet: A lentiviral plasmid for the expression of ABCG2 (pSIN4-EF2-ABCG2-IRES-Neo) was obtained from Addgene.

Techniques: Control, shRNA, Incubation, Transduction

Journal: Cellular & Molecular Biology Letters

Article Title: Limbal niche cells can reduce the angiogenic potential of cultivated oral mucosal epithelial cells

doi: 10.1186/s11658-018-0133-x

Figure Lengend Snippet: Antibodies used for immunofluorescent staining

Article Snippet: ABCG2 , BIOSS (BS-0662R) , 1:100.

Techniques:

Journal: Cellular & Molecular Biology Letters

Article Title: Limbal niche cells can reduce the angiogenic potential of cultivated oral mucosal epithelial cells

doi: 10.1186/s11658-018-0133-x

Figure Lengend Snippet: Identification and quantification of cell markers of cultivated oral mucosal epithelial cells (COMECs) co-cultured with LNCs and 3 T3 cells. a Representative images of p63α, ABCG2, Ki67, and CK3 in different culture systems. b The percentages of positive cells in different groups were approximately the same ( p > 0.05). The results were expressed as mean ± standard deviation (p63α, ABCG2, and CK3) or median with interquartile range (Ki67). NC: negative control, LNCs: limbal niche cells, ABCG2: ATP binding cassette subfamily G member 2, CK3: cytokeratin 3, scale bars: 20 μm

Article Snippet: ABCG2 , BIOSS (BS-0662R) , 1:100.

Techniques: Cell Culture, Standard Deviation, Negative Control, Binding Assay