Journal: International Journal of Biological Sciences
Article Title: Glycine Decarboxylase Regulates Renal Carcinoma Progression via Interferon Stimulated Gene Factor 3-Mediated Pathway
doi: 10.7150/ijbs.104458
Figure Lengend Snippet: GLDC regulates RCC cell progression via ISGF3 pathway. (A) Western blot of GLDC, IRF9, STAT2, STAT1, SETD2, GAPDH (loading control), H3K36me3, and H3 (loading control for H3K36me3) in indicated cells. To confirm in Caki-2 cells, we suppressed the expression of GLDC in Caki-2 cells with three shRNA constructs (shGLDC #1, shGLDC #4 and shGLDC #5), and observed the expression of indicated proteins. (B) Quantitative PCR (qPCR) analysis of the ISGs: OSA1, IFIT1, IFI44, IFI44L, CCL5, MX1, and IFI27 of indicated cells. Data are shown as the means ± SD ( n =3). (C) Western blot analysis of indicated proteins in ACHN CTL and shGLDC cells transfected with siRNA of negative control (siNC), STAT2 and/ or IRF9 in ACHN knock-downed GLDC cells compared to control (siNC)-siRNA cells. The cellular growth of indicated cells was on the right side ( n =3). (D) Colony formation of indicated cells. Quantification of staining intensity was performed at a wavelength of 570 nm ( n =3). (E) qPCR analysis of IFN-α in ACHN CTL and shGLDC treated with Poly (I:C) (10 µg) or transfected with Poly (I:C) (1 µg) for 24 h. Data are shown as the means ± SD ( n =3). p values were calculated using two-tailed unpaired Student t-test. * p < 0.05, ** p < 0.01, and *** p < 0.001. CTL, control; shGLDC, knockdown of GLDC; EV, corresponding control for OE GLDC; OE GLDC, overexpression of GLDC.
Article Snippet: Short interfering RNAs (siRNAs) for STAT2 and IRF9 came from Bioneer, Republic of Korea.
Techniques: Western Blot, Control, Expressing, shRNA, Construct, Real-time Polymerase Chain Reaction, Transfection, Negative Control, Staining, Two Tailed Test, Knockdown, Over Expression