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Fig. 5 | Cell Communication and Signaling

Fig. 5

From: Cancer-associated fibroblasts promote prostate tumor growth and progression through upregulation of cholesterol and steroid biosynthesis

Fig. 5

Effects of HMGCS2 knockdown and overexpression on PCa cell growth. a LNCaPabl cells were stably infected with a doxycycline-inducible shHMGCS2 vector (ABLshHMGCS2). Following treatment with 1 μM doxycycline, HMGCS2 was effectively downregulated on protein level compared to the mock control as shown by Western analysis. GAPDH was used as loading control. b ABLshHMGCS2 cells were seeded into 96 well plates and incubated in the absence or presence of doxycycline over 5 days. Cell viability was determined with CellTiterGlo viability assay (Promega). Representative images were taken at the end of treatment (magnification 10x). c ABLshHMGCS2 cells were seeded into ULC 96 well plates (Corning) and allowed to form spheroids over 4 days. Then, 1 μM doxycycline (dox) and 5 μM enzalutamide (enza) were added. Cell viability was determined through CellTiterGlo viability assay after 10 days of treatment. Medium was exchanged twice a week. Representative images were taken at day 10 with IncuCyte S3 software. d LNCaP cells were transiently transfected with a HMGCS2 plasmid (LNCaP_HMGCS2). HMGCS2 overexpression was confirmed 72 h afterwards by Western blotting. GAPDH was used as internal control. e LNCaP cells were transiently transfected with a HMGCS2 plasmid and seeded into a 96 well ULC plate (Corning) to allow 3D spheroid formation. After 4 days, 5 μM enzalutamide was added in RPMI with 10% CS_FCS. After 10 days, cell viability was measured via CellTiterGlo assay. Representative images were taken at the end of treatment with IncuCyte S3 software. Data represent the mean plus SEM from at least three independent experiments. (* P < 0.05, ** P < 0.01)

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