Additional file 1: of IMP1 regulates UCA1-mediated cell invasion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p

Table S1. Primers used for qPCR, RT-PCR and siRNA interference. (DOC 53 kb

Additional file 6: of IMP1 regulates UCA1-mediated cell invasion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p

Table S2. miRNAs associated with UCA1. (DOCX 13 kb

Additional file 5: of IMP1 regulates UCA1-mediated cell invasion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p

Figure S3. UCA1 is associated with IMP1 and CNOT1 and with miR-185-5p. (A) Vectors expressing UCA1 or UCA1-MS2 were transiently transfected into MDA231/IMP1-GFP cells. Pulldown assays were performed to analyze the association of IMP1 and CNOT1 with UCA1-MS2. Representative images indicate that both IMP1 and CNOT1 co-precipitated with UCA1. Control: cells transfected with MS2-untagged UCA1. (B) Putative binding site of UCA1 for miR-185-5p. (C) Interaction of miR-185-5p with UCA1-MS2 was examined in the pulldown material. Relative levels of miR-185-5p in the precipitates were statistically analyzed as means ± SD from three independent experiments: **P < 0.01 as determined by Student’s t test. (TIFF 1227 kb

Additional file 9: of IMP1 regulates UCA1-mediated cell invasion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p

Figure S6. A proposed model of IMP1 to regulate the sponge effect of UCA1 for miR-122-5p. (A) UCA1 sponges miR-122-5p, reducing miR-122-5p interaction with target mRNA. (B) Increasing IMP1 expression allows IMP1 to bind to UCA1 and to release miR-122-5p from UCA1. This increases the availability of miR-122-5p to interact with target mRNA. (C) Binding to target mRNA allows miR-122-5p to assert its posttranscriptional function. (D) IMP1 binds to UCA1 and recruits it to the CCR4-NOT1 complex, initiating UCA1 decay process. (TIFF 1116 kb

Additional file 2: of IMP1 regulates UCA1-mediated cell invasion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p

miRNAs associated with UCA1. (XLSX 375 kb

Additional file 8: of IMP1 regulates UCA1-mediated cell invasion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p

Figure S5. Effect of UCA1 on the invasive abilities of MCF7 cells. Histograms show the effect of UCA1 on the invasive abilities of MCF7 cells. Values represent the means ± SD from three independent experiments; **P < 0.01, *P < 0.05 as determined by one-way ANOVA followed by Tukey’s multiple comparison tests. (TIFF 992 kb

Additional file 7: of IMP1 regulates UCA1-mediated cell invasion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p

Figure S4. IMP1 knockdown increases the expression of miR-122-5p target mRNAs. (A) Cellular levels of miR-122-5p are not affected by IMP1-GFP expression. (B) After MS2 pulldown experiments, levels of miR-122-5p in the supernatants were analyzed by qPT-PCR. Levels of miR-122-5p were normalized to GAPDH mRNA from three independent experiments: **P < 0.01 as determined by Student’s t test. (C) RT-qPCR was applied to measure the levels of PKM2 and IGF-1R mRNAs in IMP1 knockdown T47D cells. Levels of the mRNAs were normalized to GAPDH mRNA from three independent experiments: *P < 0.05 as determined by Student’s t test. (TIFF 884 kb

The <i>gl13</i> gene encodes a putative ABC transporter G family protein.

<p>The amino acid sequence of the 2 GL13 isoforms (GRMZM2G118243_P01 and GRMZM2G118243_P02) were aligned to 9 homologs downloaded from GenBank (Table S2). Neighbor-joining analysis was used to generate a phylogenetic tree, which was bootstrapped over 1,000 cycles, using MEGA5.0. </p

Sequence analysis of 11 <i>gl13</i> alleles.

<p>The positions of detected lesions are indicated by arrows within the <i>gl13</i> gene. Green boxes indicate gene coding region (CDS) and yellow boxes indicate un-translated region (UTR). Alleles are numbered according to Table 1. Four EMS-induced premature terminal codon (PTC) mutations are shown in red; two EMS-induced non-synonymous substitutions are shown in green; an EMS-induced G->A transition is shown in blue; the location of a <i>Mu</i> insertion (belong to Mu1 family [25]) is shown in pink; the location of a C->A transversion found in two spontaneous alleles are shown in yellow; and a 2bp deletion detected in one EMS-induced allele is shown in black. </p

Phenotypic characterization of the <i>gl13-ref</i> mutant phenotype.

<p>(A) Comparisons of gross morphology and epicuticular wax accumulation and morphology in mutant (upper) and wild-type (lower) seedlings; (B) Comparisons of paraffin sections (10x) of leaves from mutant (left) and wild-type (right) seedlings stained with Fast Green; (C) Detached leaves from mutant (upper) and wild-type (lower) immediately after harvest and after 2 hours at room temperature; (D) Water loss from detached mutant and wild-type seedling leaves at room temperature. Error bars = SE. </p