Fig. 4
From: AntimiR uptake by human proximal tubule epithelial cells is predominantly by macropinocytosis
Fluorescence microscopy of human primary proximal tubule epithelial cells (PTEC) maintained on transwell inserts and treated with 1 mg/mL Texas Red-labelled dextran and either 40 nM FAM-labelled antimiR (a) or 0.1 mg/mL FITC-labelled albumin (b) for 24 h after 1 h pre-treatment with given concentration of the macropinocytosis inhibitor 5-(N-ethyl-N-isopropyl)-amiloride (EIPA) or dimethyl sulfoxide control media (0 nM EIPA). Images shown are representative of 6 images acquired at random from each condition for each biological repeat. Untreated PTEC in left column were not treated with EIPA, antimiR or dextran, as autofluorescence control. Single-channel images have been inverted for clarity. Effect of EIPA on uptake of labelled reagents used in (a) and (b) are quantified in (c) and (d) respectively as areas of FITC, FAM or Texas Red channel above threshold set by untreated controls and reported relative to 0 nM EIPA treatment. Mean ± standard deviation of (a: n = 3, b: n = 2) biological repeats are presented by bars with individual results as shapes. Statistically significant difference in EIPA treatments from 0 nM was assessed by paired t-test *P < .05. Images acquired with ZEISS AxioImager with X40 lens.
