Fig 1: The effects of miR-184 mimic on Du145 cells were rescued by DLX1 overexpression. (A) The transfection efficiency of pcDNA3.1-DLX1 in Du145 cells was tested using reverse transcription-quantitative PCR. **P<0.01 vs. pcDNA3.1. (B) The inhibitory effects of miR-84 mimics on Du145 cell viability were reversed by co-transfection with pcDNA3.1-DLX1. *P<0.05 and **P<0.01 vs. control, ##P<0.01 vs. miR-184 mimic and &P<0.05 vs. miR NC + pcDNA3.1-DLX1. (C and D) The inhibitory effects of miR-184 mimics on Du145 cell migration were reversed by co-transfection with pcDNA3.1-DLX1. (C) Representative images of the Du145 cell monolayer following transfection with miR-184 mimic or co-transfection with miR-184 mimic and pcDNA3.1-DLX1. (D) Quantified results from. (C) Scale bar, 100 µm. **P<0.01 vs. control and ##P<0.01 vs. miR-184 mimic. (E and F) The inhibitory effects of miR-184 mimic on Du145 cell invasion were reversed by co-transfection with pcDNA3.1-DLX1. (E) Representative images of invading Du145 cells following transfection with miR-184 mimic or co-transfection with miR-184 mimic and pcDNA3.1-DLX1. (F) Quantified results from. (E) Scale bar, 200 µm. **P<0.01 vs. control and ##P<0.01 vs. miR-184 mimic. DLX1, distal-less homeobox 1; miR, microRNA; OD, optical density.
Fig 2: DLX1 expression. a No correlation between expression of DLX1 and degree of methylation in human forebrains (pyrosequencing value at CpG [hg19]chr2:172,953,097) [Pearson’s correlation analysis including both PSP patients (n = 69, gray dots) and controls (n = 67, white dots)]. Expression of DLX1 did not differ between patients and controls (Welch´s corrected unpaired t-test, n.s. = not significant, bar plot with mean and SEM). b Significant correlation between expression of DLX1AS and the degree of methylation. Expression of DLX1AS is significantly reduced in patients as compared to controls (***P < 0.001, Welch´s corrected unpaired t-test, bar plot with mean and SEM). c, d No difference between the amount of DLX1 protein in white matter of frontal lobe of PSP patients and controls (co.). c Significant increase of DLX1 protein in frontal lobe gray matter of PSP as compared to controls. d (n = 8 per group, **P < 0.01, Welch´s corrected unpaired t-test, bar plot with mean and SEM). β-Actin was used as loading control. e No difference in immunoreactivity of DLX1 in white matter of gyrus frontalis between PSP and controls. f Significant increase of DLX1 protein in frontal lobe gray matter of PSP patients as compared to controls (n = 24 PSP, n = 9 controls, **P < 0.01, Mann–Whitney Test). Scale bar: 100 µm. The line in the middle of the box and whisker graph represents the median (50th percentile). The box extends from the 25th to 75th percentile. The whiskers extend from the lowest to the highest value
Fig 3: Up‐regulation of miR‐539 or silencing of DLX1 restrains PCa cell migration. A and C, PC3 cell migration and the quantitative analysis in response to miR‐539 mimic, miR‐539 inhibitor, si‐DLX1 and miR‐539 inhibitor + si‐DLX1 measured by scratch test; B and D, DU145 cell migration and the quantitative analysis in response to miR‐539 mimic, miR‐539 inhibitor, si‐DLX1 and miR‐539 inhibitor + si‐DLX1 measured by scratch test; *P < 0.05, compared with the blank group and the NC group; miR‐539, microRNA‐539; PCa, prostate cancer; NC, negative control; si, small interfering
Fig 4: PCa tissues display poor expression of miR‐539 and E‐cadherin yet high expression of DLX1, Smad4, c‐Myc, vimentin, Snail1 and SLUG. A, miR‐539 expression and mRNA expression of DLX1, Smad4, c‐Myc and vimentin, Snail1, SLUG and E‐cadherin in the PCa tissues and the adjacent normal tissues detected by RT‐qPCR; B and C, Western blot analysis of DLX1, Smad4, c‐Myc and vimentin, Snail1, SLUG and E‐cadherin proteins in the PCa tissues and the adjacent normal tissues; *P < 0.05, compared with the adjacent normal tissues; PCa, prostate cancer; miR‐539, microRNA‐539; DLX1, distal‐less homeobox 1; RT‐qPCR, reverse transcription quantitative polymerase chain reaction
Fig 5: DLX1 is a direct target of miR-184. (A) Sequence of the miR-184-binding site in DLX1 3'UTR. (B) Relative luciferase activity in Du145 cells co-transfected with miR-184 mimic or miR-NC and DLX1-3'UTR (wt) or DLX1-3'UTR (mut) and corresponding quantification. **P<0.01 vs. miR-NC. (C) Pearson's analysis between miR-184 and DLX1 expression based on the PC tissue data from TCGA database. P<0.0001. (D) Reverse transcription-quantitative PCR measurement of DLX1 expression following miR-184 overexpression, *P<0.05. (E and F) Expression of DLX1 protein was measured in Du145 cells using western blot analysis following transfection with miR-184 mimic or miR-NC; the results were normalized to GAPDH. (E) Representative western blot images showing DLX1 and GAPDH expression. (F) Quantified data from. (E) *P<0.05. DLX1, distal-less homeobox 1; miR, microRNA; 3'UTR, 3' untranslated region; wt, wild-type; mut, mutant; NC, negative control.
Supplier Page from Abcam for Anti-Dlx1 antibody