Fig 1: Knockdown of PRRX1 enhanced the inhibitive effects of gemcitabine on tumor growth in vivo. (A) The size, volume, and weight of tumors were detected in the shNC, sh1#PRRX1, gemcitabine + shNC, and gemcitabine + sh1#PRRX1 groups. ***, P<0.001 compared with shNC, ###, P<0.001 compared with gemcitabine + sh1#PRRX1. (B) The expression of PRRX1, Ki67, and Beclin-1 was examined by immunohistochemistry. PRRX1, paired related homeobox 1; NC, negative control.
Fig 2: Schematic diagram. PRRX1 cooperates with FOXM1 to regulate the expressions of the autophagy proteins (LC3 and Beclin-1), and reduces gemcitabine-induced cytotoxicity in BC cells. PRRX1, paired related homeobox 1; FOXM1, Forkhead box M1; BC, bladder cancer.
Fig 3: PRRX1 was up-regulated in BC cells and enhanced cell viability. (A) The protein expression of PRRX1 was detected using western blot. *, P<0.05, **, P<0.01, ***, P<0.001 compared with HCV-29 cells. (B) The overexpression and knockdown efficiency of PRRX1 were verified using western blot. ***, P<0.001 compared with Vector, ###, P<0.001 compared with shNC. (C) Cell viability after overexpressing or suppressing PRRX1 was detected by MTT assay. *, P<0.05, **, P<0.01 compared with Vector. ##, P<0.01, ###, P<0.001 compared with shNC. (D) Cell apoptosis after PRRX1 up-regulation or repression was examined using flow cytometry. **, P<0.01 compared with Vector, ##, P<0.01 compared with shNC. BC, bladder cancer; PRRX1, paired related homeobox 1; NC, negative control; OD, optical density; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide.
Fig 4: PRRX1 was highly expressed in BC tissues. (A) The mRNA expression of PRRX1 was verified in normal and tumor tissues using real-time quantitative PCR. **, P<0.01 compared with normal. (B) The overall survival rate of patients with PRRX1. Black line: BC patients with low PRRX1 expression; Green line: BC patients with high PRRX1 expression. (C) The protein expression of PRRX1 in BC tissues was detected by immunohistochemistry. (D) The protein expression of PRRX1 in BC tissues was examined using western blot. BC, bladder cancer; PRRX1, paired related homeobox 1; PCR, polymerase chain reaction.
Fig 5: PRRX1 weakened gemcitabine-induced cytotoxicity and induced autophagy in BC cells. (A) Cell viability was determined after PRRX1 overexpression or repression in BC cells treated with different doses of gemcitabine by conducting an MTT assay. (B) Cell apoptosis was measured after overexpressing or repressing PRRX1 in BC cells treated with gemcitabine (1 µM) by flow cytometry. **, P<0.01 compared with Control, &, P<0.05 compared with Gemcitabine. ##, P<0.01 compared with Control, $$, P<0.01 compared with Gemcitabine. (C) The autophagosome puncta of GFP-LC3 were examined in in BC cells treated with gemcitabine (1 µM) by immunofluorescence assay. *, P<0.05 compared with Control, &&, P<0.01 compared with Gemcitabine. ##, P<0.01 compared with Control, $$, P<0.01 compared with Gemcitabine. Scale: 25 μm. (D) The protein expression of LC3II/I was measured using western blot. ***, P<0.001 compared with Control, &&&, P<0.001 compared with Gemcitabine. ###, P<0.001 compared with Control, $$$, P<0.001 compared with Gemcitabine. BC, bladder cancer; PRRX1, Paired related homeobox 1; NC, negative control; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide.
Supplier Page from Abcam for Anti-PRX-1 antibody