Fig. 5

Evaluation of autophagy levels and flux analysis following miR-193b overexpression in KYSE450 oesophageal cancer cells. a Western blot analysis of LC3 II (autophagy marker) expression at 48 and 72 h post-transfection with miR-193b mimic or negative control mimic (5 nM). β-actin was used as a loading control. LC3 II levels were normalised to actin and quantification is shown in the lower panel. Data is presented as mean +/- S.E.M (n = 2). b KYSE450 cells were transfected with negative control mimic or miR-193b mimic (5 nM). Twenty-four hours following transfection, cells were treated with chloroquine (10 μM) and cells were assessed for autophagy induction 48 h after treatment using the Cyto-ID autophagy detection assay. (i) Representative FACS analysis of Cyto-ID levels of untreated, chloroquine treated, negative control mimic and miR-193b transfected cells. (ii) Representative FACS analysis of chloroquine, negative control mimic and miR-193b mimic transfected cells treated with chloroquine. The right hand panel shows the corresponding mean fluorescence intensity of all treatments. Asterisks indicate statistical significance determined by t-test (*p < 0.05)