Figure 1
From: NADPH oxidase NOX4 is a glycolytic regulator through mROS-HIF1α axis in thyroid carcinomas
NOX4 Knockdown results in inhibition of TPC-1 Proliferation. (A,B) Transcriptional expression of NOX4 in TPC-1 cells after 48 hours treated with lentiviral transduction particles targeting NOX4 mRNA (A). Protein expression level of NOX4 after 72 hours treated with lentiviral transduction particles targeting NOX4 mRNA (B). Con for shNOX4 control lentivirus, #1 for shNOX4#1 lentivirus, and #2 for shNOX4#2 lentivirus. **P < 0.01. (C) Viability assay for TPC-1 cells expressing shControl or shRNA against NOX4 (shNOX4#1,#2) which were cultured in normoxia (21% O2) and hypoxia (1% O2) respectively for 48 hours using CCK8 assay (n = 8). **P < 0.01. (D,E) Western blot for normoxia (21% O2) and hypoxia (1% O2) in TPC-1 cell clones after infected with either shNOX4 control lentivirus and shNOX4#1and shNOX4#2 lentivirus (D). The blots were quantified using ImageJ software (n = 3). **P < 0.01. (F,G) TPC-1 cells transduced with shNOX4 control or two NOX4-directed shRNAs were injected subcutaneously in the flanks of nude mice. Tumor growth was quantified with a caliper at the indicated time intervals for 20 days (F). After the measurement, these mice were euthanized and then stripped of the subcutaneous transplantation tumor to take pictures at 20 days (G). Data were analyzed using the two-sided unpaired Student’s t test. Mean ± SEM. **p < 0.01.
