Journal: Bioscience Reports

Article Title: miR-484 is associated with disease recurrence and promotes migration in prostate cancer

doi: 10.1042/BSR20191028

Figure Lengend Snippet: Selected candidate gene targets of miR-484 in DU145 and PC-3

Article Snippet: Primary antibodies used were PSMG1 (1:1000 of Cat# 13378) from Cell Signaling (Danvers, MA) and β-Actin (1:5000 of sc-47778) from Santa Cruz (Dallas, TX).

Techniques:

Journal: Bioscience Reports

Article Title: miR-484 is associated with disease recurrence and promotes migration in prostate cancer

doi: 10.1042/BSR20191028

Figure Lengend Snippet: ( A ) Sequence-verified wild-type (WT) 3′ UTR or mutant (MUT) 3′ UTR of PSMG1 was cloned into a luciferase reporter plasmid and transfected into DU145 human prostate cancer cells with Renilla plasmid (for normalization) and with either negative control miR or miR-484 mimic. Y -axis indicates the ratio of luciferase to Renilla , relative to the miRNA mimic, when negative control was set to 1.0. N = 4 with error bars (± S.D.), and * P <0.05 using unpaired t test with Welch’s correction. ( B ) Transfection of DU145 cells with miR-484 mimic reduces PSMG1 protein expression. Shown are Western blot results with β-actin expression as a reference and loading control.

Article Snippet: Primary antibodies used were PSMG1 (1:1000 of Cat# 13378) from Cell Signaling (Danvers, MA) and β-Actin (1:5000 of sc-47778) from Santa Cruz (Dallas, TX).

Techniques: Sequencing, Mutagenesis, Clone Assay, Luciferase, Plasmid Preparation, Transfection, Negative Control, Expressing, Western Blot, Control

Journal: Bioscience Reports

Article Title: miR-484 is associated with disease recurrence and promotes migration in prostate cancer

doi: 10.1042/BSR20191028

Figure Lengend Snippet: ( A ) miR-484 inhibition had no effect on the viability of 22Rv1, DU145 or PC-3 cells over 5 days. Cell viability was measured using fluorescence ( E x/ E m = 544/590 nm). Differences between miR-484 inhibitor and negative control miR were non-significant (NS) according to unpaired t test with Welch’s correction; N = 3 with error bars (± S.D.). ( B ) miR-484 inhibition had no effect on apoptosis of 22Rv1, DU145 or PC-3 cells. Apoptosis was measured 48 h after miR transfection using caspase-3 activity as a readout, which was measured using fluorescence ( E x/ E m = 485/520 nm); N = 3 with error bars (± S.D.). Differences between miR-484 inhibitor and negative control miR were NS according to unpaired t test with Welch’s correction. In contrast, miR-484 inhibition reduces migration ( C ) and invasion ( D ), which is rescued by inhibition of PSMG1 with siRNA ( E and F ). Y -axis denotes % migration or invasion relative to negative control at 48 h, set at 100%; N = 3 with error bars (± S.D.), and * P <0.05 using unpaired t test with Welch’s correction; Scr, scrambled.

Article Snippet: Primary antibodies used were PSMG1 (1:1000 of Cat# 13378) from Cell Signaling (Danvers, MA) and β-Actin (1:5000 of sc-47778) from Santa Cruz (Dallas, TX).

Techniques: Inhibition, Fluorescence, Negative Control, Transfection, Activity Assay, Migration