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Fig. 6 | Breast Cancer Research

Fig. 6

From: ELF5 isoform expression is tissue-specific and significantly altered in cancer

Fig. 6

Alterations in cell line E74-like factor 5 (ELF5) isoform levels result in a similar phenotype, characterized by decreased cell number, decreased estrogen-related proteins, and nuclear localization. a and c Time course of T47D (a) and MDA-MB-231 (c) pHUSH clonal cell line growth with and without doxycycline (Dox) over 5 days. Graphs show the natural logarithm (Ln) of spectrophotometric assay absorbance value (y-axis) plotted against day (x-axis). p values compare minus Dox and plus Dox slopes for each cell line. One experiment is shown. b Representative light microscopic images of T47D cells with and without doxycycline, taken at day 4. d Western blots for estrogen-related proteins from T47D time courses, days 0–5. e Western blots for estrogen-related proteins from MDA-MB-231 time courses, days 2–5. f Quantitative polymerase chain reaction for ELF5 (day 5 time course samples) minus and plus Dox. g Western blots for V5 at days 4 and 5 minus and plus Dox, 65 μg per lane (T47D-ELF5-isoform 2-V5 line) or 25 μg per lane (all others). Bottom panel shows representative samples from MDA-MB-231 cell lines, demonstrating size difference between isoforms 2 and 3. h Immunofluorescent images of MDA-MB-231-ELF5-isoform 2-V5 clone 7 cells. Blue = nuclei (4′,6-diamidino-2-phenylindole [DAPI]), red = V5, green = hemagglutinin (HA). Arrows mark cells with double-isoform 2 and 3 expression

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