Fig. 3: Hypoxia induces glutaminase 1 (GLS1) expression in a HIF-1α-dependent manner in colorectal cancer cell lines. | Cell Death & Disease

Fig. 3: Hypoxia induces glutaminase 1 (GLS1) expression in a HIF-1α-dependent manner in colorectal cancer cell lines.

From: Glutaminase 1 expression in colorectal cancer cells is induced by hypoxia and required for tumor growth, invasion, and metastatic colonization

Fig. 3

a GLS1 protein expression in colorectal cancer cell lines. b Reverse transcription and quantitative real-time PCR (RT-qPCR) was performed to quantify GLS1 mRNA levels in colorectal cancer cell lines (HT29 and Caco2) following exposure to 20 or 1% O2 for 24 h. For each sample, the expression of GLS1 mRNA was quantified relative to 18S rRNA and then normalized to the result obtained from cells at 20% O2. Statistical analysis was performed before normalization. Data are shown as mean ± SEM; n = 3. **p < 0.01, ***p < 0.001 vs. 20% O2 (Student’s t-test). c Immunoblot assays were performed to analyze GLS1 and HIF-1α protein expression in HT29 and Caco2 cell lines following exposure to 20 or 1% O2 for 48 h. d levels of GLS1 mRNA were analyzed by RT-qPCR in HT29 subclones, which were stably transfected with non-targeting control (NTC) or vector encoding hypoxia-inducible factor (HIF)-1α short hairpin RNA (shRNA) (sh1α-1 or sh1α-2), and exposed to 20 or 1% O2 for 24 h (mean ± SEM; n = 3). *p < 0.05, ***p < 0.001 vs. NTC at 20% O2; ###p < 0.001 vs. NTC at 1% O2 (analysis of variance (ANOVA) with Bonferroni post-test). e Immunoblot assays were conducted using lysates prepared from HT29 subclones exposed to 20 or 1% O2 for 48 h

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