The GATA factor HANABA TARANU promotes runner formation by regulating axillary bud initiation and outgrowth in cultivated strawberry

A runner, as an elongated branch, develops from the axillary bud (AXB) in the leaf axil and is crucial for the clonal propagation of cultivated strawberry (Fragaria x ananassa Duch.). Runner formation occurs in at least two steps: AXB initiation and AXB outgrowth. HANABA TARANU (HAN ) encodes a GATA transcription factor that affects AXB initiation in Arabidopsis and promotes branching in grass species, but the underlying mechanism is largely unknown. Here, the function of a strawberry HAN homolog FaHAN in runner formation was characterized. FaHAN transcripts can be detected in the leaf axils. Overexpression (OE) of FaHAN increased the number of runners, mainly by enhancing AXB outgrowth, in strawberry. The expression of the strawberry homolog of BRANCHED1 , a key inhibitor of AXB outgrowth in many plant species, was significantly downregulated in the AXBs of FaHAN -OE lines, whereas the expression of the strawberry homolog of SHOOT MERISTEMLESS, a marker gene for AXB initiation in Arabidopsis, was upregulated. Moreover, several genes of gibberellin biosynthesis and cytokinin signaling pathways were activated, whereas the auxin response pathway genes were repressed. Further assays indicated that FaHAN could be directly activated by FaNAC2, the overexpression of which in strawberry also increased the number of runners. The silencing of FaNAC2 or FaHAN inhibited AXB initiation and led to a higher proportion of dormant AXBs, confirming their roles in the control of runner formation. Taken together, our results revealed a FaNAC2-FaHAN pathway in the control of runner formation and have provided a means to enhance the vegetative propagation of cultivated strawberry.Peer reviewe

Flot-2 Expression Correlates with EGFR Levels and Poor Prognosis in Surgically Resected Non-Small Cell Lung Cancer.

We previously reported that expression of Flotillin 2 (Flot-2), a protein isolated from caveolae/lipid raft domains, increased significantly in nasopharyngeal carcinoma (NPC) compared with normal tissues. Signal transduction through epidermal growth factor receptors (EGFR) and Flot-2 play an important role in cancer development, but their precise role in lung cancer has not been investigated. In this study, we have investigated the correlation between the expression of Flot-2 and EGFR, which increase significantly in non-small cell lung cancer (NSCLC) patients (n=352) compared with non-cancer tissues. Additionally, patients with advanced stages of NSCLC had higher positive expression of Flot-2 and EGFR than patients with early stages. NSCLC patients with increased expression of Flot-2 and EGFR had significantly less overall survival rates than patients with less expression of Flot-2 and EGFR. Taken together, our data suggest that increased expression of Flot-2 and EGFR in NSCLC patients is inversely proportional to the disease prognosis and that increased expression of Flot-2 associated with increased EGFR may serve as a biomarker to predict poor disease prognosis

Kaplan-Meier cures for overall survival of lung SCC and ADC patients with expression of Flot-2 and EGFR.

<p>Kaplan-Meier analysis was used to plot the overall survival curves of 159 cases of lung SCC and 193 cases of lung ADC patients with differential expression of Flot-2, EGFR, and combined expression of either of these two proteins, which statistical significance was assessed by log-rank test. (A) Lung SCC patients with positive expression of Flot-2 protein showed worse overall survival rates compared to patients with negative Flot-2 (P = 0.043, two sided). (B) Positive expression of EGFR had no significantly correlation with overall survival rates of lung SCC patients (P> 0.05, two sided). (C) Kaplan-Meier curves showed lung SCC patients with positive expression with either of Flot-2 and EGFR proteins had worse overall survival rates than these with all negative staining of two proteins above (P = 0.02, two sided). (D) Lung ADC patients with positive expression of Flot-2 had worse overall survival rates than that with negative one (P = 0.007, two sided, respectively). (E) Lung ADC patients with positive expression of EGFR had worse overall survival rates than that with negative one (P = 0.033, two sided, respectively). (F) Lung ADC patients with positive expression with either of Flot-2 and EGFR proteins showed worse overall survival rates compared with all negative staining of two proteins above (P = 0.005, two sided).</p

Expression of Flot-2 and EGFR proteins in lung SCC and lung ADC compared to the control of non-cancerous lung tissues.

<p>Results showed that there were significant differences between the groups which were statistically evaluated by chi-square test.</p

352 cases of non small cell lung cancer (NSCLC) patient’s characteristics.

<p>352 cases of non small cell lung cancer (NSCLC) patient’s characteristics.</p

Expression of Flot-2 protein in lung SCC cells, lung ADC cells and control of non-cancerous lung tissues were detected by IHC using specific antibody as described in the section of materials and methods.

<p>Strong positive staining of Flot-2 protein was found on cell membranes of lung SCC and lung ADC cells (Fig 1A and 1B, 20×, IHC, DAB staining). Negative staining of Flot-2 was showed in non-cancerous lung tissue (Fig 1C, 20×, IHC, DAB staining). Negative control showed no Flot-2 staining in lung ADC cells (Fig 1D, 20×, IHC, DAB staining).</p

Expression of EGFR protein in lung SCC cells, lung ADC cells and the control of non-cancerous lung tissue were detected by IHC using specific antibody as described in the section of materials and methods.

<p>Strong positive staining of EGFR protein was found in cell membranes and cytoplasm of lung SCC and lung ADC cells (Fig 2A and 2B, 20×, IHC, DAB staining). Negative staining of EGFR was showed in non-cancerous lung tissue (Fig 2C, 20×, IHC, DAB staining). Negative control showed no EGFR staining in the lung SCC cells (Fig 2D, 20×, IHC, DAB staining).</p

Additional file 1: of Increased expression of Cks1 protein is associated with lymph node metastasis and poor prognosis in nasopharyngeal carcinoma

The original data of IHC scoring. The expression of CKS1 and p27KIP1 had been tested in 168 cases of paraffin-embedded tissue from the primary NPC patients and 49 cases of non-cancerous nasopharyngeal control specimen from independent patients by IHC. The percentage of positive cells was divided into five grades (percentage scores):<10% (0), 11–25% (1), 26–50% (2), 51–75%(3), and >75% (4). The intensity of staining was divided into four grades (intensity scores): no staining (0), light brown (1), brown (2),and dark brown (3). Staining positivity was determined by the formula: overall scores = percentage score × intensity score. The total score ranged from 0 to 12, with negative staining (0–1) and positive expression (2–12). At last, 0 presented negative staining and 1 presented positive staining in the data file. (XLS 42 kb