Journal: Nucleic Acids Research

Article Title: Periodic-shRNA molecules are capable of gene silencing, cytotoxicity and innate immune activation in cancer cells

doi: 10.1093/nar/gkv1488

Figure Lengend Snippet: Cytotoxicity and gene knockdown studies with p-shRNA in different cell lines. ( A ) Cell viability dose-response curves for p-shRNA, RNase T1-digested p-shRNA and siRNA delivered with Lipofectamine to HeLa cells. Viability was measured by the CellTiterGlo assay and normalized to untreated cells. p-shRNA for this experiment was derived from template 6. ( B ) IC 50 values ± 95% C.I. (based on three parameter log-logistic model) calculated from viability curves for cell lines treated with p-shRNA (I = p-shRNA-21, II = p-shRNA-25; derived from templates 17 and 18) or HMW poly-I:C complexed with Lipofectamine. ( C ) Cell viability dose-response curves for p-shRNA-25 and poly-I:C delivered with either Lipofectamine or TransIT-X2 to SKOV3 cells. ( D ) Caspase activation measured using CaspaseGlo luminescence assays 14 h after treating SKOV3 cells with p-shRNA-25- or poly-I:C-Lipofectamine complexes at 300 ng/ml. Values correspond to the means of three biological replicates (±s.e.m.). ( E ) Mean GFP expression determined by flow cytometry was measured in HeLa cells 72 h following transfection with 10 nM p-shRNA (p-shGFP-21 or p-shLUC-21) complexed with TransIT-X2. Values were derived from mean fluorescence and correspond to the mean of three biological replicates (±s.e.m.) relative to untreated cells. ( F ) Luciferase expression was measured as bioluminescence normalized to total protein in SKOV3 and UCI101 cells following treatment with 15 nM p-shRNA complexed with TransIT-X2 (p-shLUC and p-shGFP used in this experiment were synthesized from templates 19 and 20). Values are presented as means for three biological replicates (±s.e.m.) relative to untreated cells. For all graphs: P < 0.05 (*), P < 0.01 (**), P < 0.001 (***).

Article Snippet: Green fluorescent protein (GFP)-expressing HeLa and A549 cell lines were purchased form CellBioLabs and maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin.

Techniques: Knockdown, shRNA, Derivative Assay, Activation Assay, Expressing, Flow Cytometry, Transfection, Fluorescence, Luciferase, Synthesized

Journal: Scientific Reports

Article Title: Freeform micropatterning of living cells into cell culture medium using direct inkjet printing

doi: 10.1038/s41598-017-14726-w

Figure Lengend Snippet: Demonstration of various cell patterns that were directly printed into culture medium. ( a ) Printed cell migration assay model with 1-mm and 0.5-mm gaps. Images were captured at 2 hours and 48 hours after printing. Scale bar: 500 µm. ( b–e ) Fluorescent images of red-, green-, and blue-labeled cell patterns. Cells were pre-labeled and printed in patterns. Scale bar: 1 mm. ( f ) Printed zigzag pattern using a heterotypic co-culture model of NIH3T3 and HEK293A cells. Scale bar: 1 mm.

Article Snippet: Mouse NIH3T3 fibroblasts and HEK293A human embryonic kidney cells were provided by Prof. Kyungtae Kim, POSTECH, Korea.

Techniques: Cell Migration Assay, Labeling, Co-Culture Assay