Fig. 3
From: NDRG1 regulates neutral lipid metabolism in breast cancer cells
NDRG1 is required for normal breast cancer cell proliferation, viability and morphology. a Immunoblots of NDRG1 and GAPDH loading control after selection of four stable lentiviral transduced shRNA expressing cell lines. A green fluorescent protein (GFP) targeting shRNA serves as negative control, and silencing was verified at 1% O2. b Cell proliferation assays: cell number was compared by comparing NDRG1 silenced cell numbers to control at the times indicated: ***p < 0.0001, N = 3/group - see Additional file 1: Figure S5. c Cleaved caspase 3 (CC3) immunofluorescence analysis of stable shRNA expressing SKBR3 cells at 1 week post selection. CC3 positive area was normalized to total cell area and each population compared: shNDRG1_1 p = 2.5 × 10− 6, shNDRG1_2 p = 1.2 × 10− 4, scale = 100 μm, N = 12/group. d Individual frames from a 48 h live cell microscopy analysis of SKBR3 cells expressing the indicated shRNAs: N = 3/group, see Additional files 3, 4, 5, 6, 7 and 8: Movies S1–S6. e Mitotic cell fractions comparing the percentage of histone H3 phsopho-serine10 positive cells in the indicated shRNA expressing cell lines: shNDRG1_1 p = 0.02, shNDRG1_2 p = 0.06, N = 12/group. f Comparison of individual cell two-dimensional area based on phalloidin stain (p < 0.001, Mann-Whitney U test, N = > 650 cells each group). Log2 two-dimensional area is shown. All bar graphs represent means +/− SD. Comparisons were by two-sided Student’s t test unless indicated otherwise