EMBO/soda-nel huggingface dataset:
Nuclear Receptor Corepressor 1 Represses Cardiac Hypertrophy (2019)
Figure 1

Figure 1. Deficiency of cardiomyocyte NCoR1 leads to cardiac hypertrophy and dysfunction. (A) Ventricular weight to tibia length ratio (VW/TL) of littermate control (LC) or cardiomyocyte-specific NCoR1 knockout (CMNKO) mice. n=10:9:12:7. (B) Ventricular weight to body weight ratio (VW/BW) of LC or CMNKO mice. n=10:8:12:7. (C) Representative H&E staining of cross sections of left ventricles. Scale bar: 50μm. (D) Representative wheat germ agglutinin (WGA) staining of cross section of left ventricles. Scale bar: 10μm. (E) Quantification of cardiomyocyte size. n=11:10:8:8. (F) qRT-PCR analysis of hypertrophy-related genes in left ventricles. n=9:9:11:8. (G) Ejection fraction and (H) Fractional shortening measured by echocardiography. n=9:7:14:10. Student's t test was used for statistical analysis.
Nuclear Receptor Corepressor 1 Represses Cardiac Hypertrophy (2019)
Figure 3

Figure 3. NCoR1 inhibits phenylephrine-induced cardiomyocyte hypertrophy. (A) Western blotting analysis of NCoR1 in neonatal rat ventricular myocytes (NRVMs) transfected with siControl or siNCoR1 for 3 days. siControl indicates control siRNA; siNCoR1, NCoR1 siRNA. (B) Representative immunofluorescence staining of α-Actinin in NRVMs transfected with siRNA for 48h and then treated with vehicle (H2O) or phenylephrine (PE) for another 48h. Scale bar: 50μm. (C) Quantification of the surface area of α-Actinin-positive NRVMs with or without NCoR1 knockdown. A total of 40 NRVMs was randomly chosen from 4 replicate coverslips for each group and used for statistical analysis. (D) qRT-PCR analysis of Acta1 and Nppa in NRVMs with or without NCoR1 knockdown. n=4:4. (E) Western blotting analysis of NCoR1-flag in NRVMs infected by control lentivirus (Control) or NCoR1-flag lentivirus (NCoR1-OV) for 4 days. (F) Representative immunofluorescence staining of α-Actinin in NRVMs infected with lentivirus for 48h and then treated with vehicle or PE for another 48h. Scale bar: 50μm. (G) Quantification of the surface area of α-Actinin-positive NRVMs with or without NCoR1 overexpression. A total of 30 NRVMs was randomly chosen from 3 replicate coverslips for each group and used for statistical analysis. (H) qRT-PCR analysis of Acta1 and Nppa in NRVMs with or without NCoR1 overexpression. n=4:3. Data represent three independent experiments. Two-way ANOVA followed by Bonferoni post-tests was used for statistical analysis.
Nuclear Receptor Corepressor 1 Represses Cardiac Hypertrophy (2019)
Figure 4

Figure 4. MEF2a and MEF2d mediate the impacts of NCoR1 knockdown on PE-induced cardiomyocyte hypertrophy. (A) Representative immunofluorescence staining of α-Actinin in NRVMs transfected with siRNA for 48h and then treated with vehicle (H2O) or PE for another 48h. Scale bar: 50μm. siMEF2a indicates MEF2a siRNA. (B) Quantification of the surface area of α-Actinin-positive NRVMs with or without knockdown of NCoR1 and/or MEF2a. A total of 15-20 NRVMs was randomly chosen from 3 replicate coverslips for each group and used for statistical analysis. (C) Representative immunofluorescence staining of α-Actinin in NRVMs transfected with siRNA for 48h and then treated with vehicle or PE for another 48h. Scale bar: 50μm. siMEF2d indicates MEF2d siRNA. (D) Quantification of the surface area of α-Actinin-positive NRVMs with or without knockdown of NCoR1 and/or MEF2d. A total of 15-19 NRVMs was randomly chosen from 3 replicate coverslips for each group and used for statistical analysis. (E) qRT-PCR analysis of Acta1 and Nppa in NRVMs. n=4. Data represent three independent experiments. Student's t test was used for statistical analysis.
Nuclear Receptor Corepressor 1 Represses Cardiac Hypertrophy (2019)
Figure 5

Figure 5. NCoR1 directly interacts with MEF2a to suppress its transcriptional activity. (A) Luciferase assays of Acta1 and Nppa promoters in HEK293FT cells transfected with NCoR1-Flag and/or MEF2a-HA or with empty plasmids. (B) Co-immunoprecipitation (Co-IP) analysis of NCoR1 and MEF2a in HEK293FT cells transfected with full-length NCoR1-Flag and MEF2a-HA. (C) Co-IP analysis of NCoR1 and truncated MEF2a in HEK293FT cells. Schematic illustration of MEF2a-HA construct is shown above the Co-IP results. (D) Co-IP analysis of MEF2a and truncated NCoR1 in HEK293FT cells. Schematic illustration of NCoR1-Flag construct is shown above the Co-IP results. (E) Luciferase assays of Acta1 and Nppa promoters in HEK293FT cells transfected with full-length MEF2a and domain-specific NCoR1. (F) Western blotting analysis of NCoR1-flag in NRVMs infected by control lentivirus (Control) or NCoR1 (1939-2453)-flag lentivirus [NCoR1(1939-2453)-OV] for 4 days. (G) Representative immunofluorescence staining of α-Actinin in NRVMs infected with lentivirus for 48h and then treated with vehicle (DMSO) or PE for another 48h. Scale bar: 50μm. (H) Quantification of the surface area of α-Actinin-positive NRVMs. A total of 30 NRVMs was randomly chosen from 3 replicate coverslips for each group and used for statistical analysis. (I) qRT-PCR analysis of Acta1 and Nppa in NRVMs. n=4:4. Data represent three independent experiments. Student's t test was used for statistical analysis in A and E. Two-way ANOVA followed by Bonferoni post-tests was used for statistical analysis in H and I.
Nuclear Receptor Corepressor 1 Represses Cardiac Hypertrophy (2019)
Figure 6

Figure 6. NCoR1 forms a complex with MEF2 and HDAC4 to repress the expression of hypertrophy-related genes in vivo. (A) ChIP analysis showing enrichment of NCoR1 in MEF2 binding regions on the promoters of Acta1 and Nppa in ventricular samples. (B) ChIP analysis of enrichment of MEF2a on promoters of Acta1 and Nppa in ventricular samples. n=3:3. (C-F) ChIP analysis of enrichment of HDAC4 (C), HDAC5 (D), acetylated-Histone 4 (ac-H4) (E), and ac-H3 (F) on promoters of Acta1 and Nppa in ventricular samples. n=4:4 for (C), n=3:3 for (D, E, F). (G) ChIP analysis of RNA Polymerase II enrichment on the transcriptional regions of Acta1 and Nppa in ventricular samples. n=3:3. (H) Working model of NCoR1 function in cardiomyocytes. TSS indicates transcription start site. Student's t test was used for statistical analysis.
Nuclear Receptor Corepressor 1 Represses Cardiac Hypertrophy (2019)
Figure 7

Figure 7. Overexpression of RIDs of NCoR1 inhibits AAC-induced cardiac hypertrophy. (A) Western blotting analysis of RIDs of NCoR1 in left ventricular samples from mice infected with AAV9 GFP or AAV9 NCoR1-RIDs (1939-2453)-Flag. (B) VW/BW of mice 2 weeks after sham operation or AAC. n=7:7:12:12. (C) Representative H&E staining of cross sections of left ventricles. Scale bar: 10μm. (D) Representative WGA staining. Scale bar: 10μm. (E) Quantification of cardiomyocyte size. n=6:6:7:8. (F) qRT-PCR analysis of hypertrophy-related genes in left ventricles. n=8:8:11:11. (G) Representative picrosirius red staining of cross sections of left ventricles. Scale bar: 20μm. (H) Quantification of fibrotic areas. n=5:5:7:7. (I) qRT-PCR analysis of fibrosis-related genes in left ventricles. n=8:8:11:11. (J) Ejection fraction and (K) Fractional shortening measured before, 2 weeks and 4 weeks after AAC. n=6:6. Two-way ANOVA followed by Bonferoni post-tests was used for statistical analysis.
Autophagic control of listeria through intracellular innate immune recognition in drosophila (2008)
figf1

(a,b) Survival of wild-type (Oregon R), yw, PGRP-LE112, PGRP-LC7454 and PGRP-LE112,PGRP-LC7454 flies after injection of wild-type L. monocytogenes (a) or Δhly L. monocytogenes (b) at 28 °C. (c) Survival of wild-type, yw and PGRP-LE112 flies, as well as flies treated with RNAi targeting PGRP-LE with the hml-Gal4 driver (hml-Gal4>PGRP-LE RNAi) and PGRP-LE112 flies with hml-Gal4–driven PGRP-LE expression (PGRP-LE112,hml-Gal4>PGRP-LE), after injection of wild-type L. monocytogenes at 28 °C. NS, not significant; *, P 0.01 (Wilcoxon-Mann-Whitney test): P = 0.0006, 0.0043 or 0.0043, for yw versus PGRP-LE112, PGRP-LC7454 or PGRP-LE112,PGRP-LC7454, respectively (a); P = 0.0095, for yw versus RNAi targeting PGRP-LE (c). Data represent the average of four independent experiments with over 30 flies of each genotype examined at the same time.
Autophagic control of listeria through intracellular innate immune recognition in drosophila (2008)
figf2

(a) Microscopy of hemocytes from third instar larvae cultured ex vivo and infected with wild-type L. monocytogenes. Hemocyte nuclei (filled arrowhead) and DNA of L. monocytogenes (open arrowhead) are visualized by DAPI staining (blue or white); the actin cytoskeleton is stained with rhodamine-labeled phalloidin (red). hml-Gal4>Atg5IR, expression of an inverted repeat sequence of Atg5 to induce RNAi against Atg5. Scale bars, 10 μm. (b) Intracellular wild-type L. monocytogenes (WT Lm) or Δhly L. monocytogenes (Δhly Lm) counted manually in DAPI-stained infected hemocytes. LE112,hml>LE, hml-Gal4–driven expression of PGRP-LE in PGRP-LE112 hemocytes; imd1, imd; RelE20, RelishE20; hml>Atg5IR, inverted repeat sequence described in a; w, hml>Atg1, hml-Gal4–driven expression of Atg1 in w hemocytes; LE112,hml>Atg1, hml-Gal4–driven expression of Atg1 in PGRP-LE112 hemocytes; PGRP-LE112+rap, PGRP-LE112 hemocytes plus rapamycin. *, P 0.001, compared with wild type (t-test). Data are representative of four experiments (a) or at least three independent experiments (b; error bars, s.d. of triplicate measurements).
Autophagic control of listeria through intracellular innate immune recognition in drosophila (2008)
figf3

Survival of yw flies, PGRP-LE112 flies, flies treated with Atg5-directed RNAi (hml-Gal4>Atg5IR) and control flies carrying hml-Gal4 (hml-Gal4>GFP), after injection of wild-type L. monocytogenes (a), Δhly L. monocytogenes (b) or E. carotovora (c) into adult flies. *, P 0.01 (Wilcoxon-Mann-Whitney test): P = 0.0024, Atg5-directed RNAi versus control (a), P = 0.0007, PGRP-LE112 versus control (a); P = 0.0055, PGRP-LC7454 versus yw (c). Data represent the average of four independent experiments.
Autophagic control of listeria through intracellular innate immune recognition in drosophila (2008)
figf4

(a) Growth of L. monocytogenes in S2 cells (S2) or S2 cells expressing PGRP-LE (S2-LE) transfected with double-stranded RNA (RNAi) specific for Atg5, Rel or Dif and dl (Dif-dl) and infected for 1.5 h with wild-type or Δhly L. monocytogenes, followed by 6 h of incubation in medium containing CuSO4 and gentamicin, quantified by plate assay of colony-forming units. *, P 0.001 (t-test). Data are representative of two independent experiments (error bars, s.d.). (b,c) Fluorescence confocal microscopy of the localization of PGRP-LE together with wild-type L. monocytogenes in S2 cells engineered to express YFP–PGRP-LE (YFP-LE) and infected for 0.5 h with wild-type L. monocytogenes (b) or Δhly L. monocytogenes (c), followed by 1 h of incubation in gentamicin-containing medium, then DAPI staining. Merge, YFP–PGRP-LE (green) and DAPI (magenta). Filled arrowheads indicate L. monocytogenes; open arrowhead indicates accumulation of YFP–PGRP-LE around the bacteria. Scale bars, 5 μm. Data are representative of three experiments.
Autophagic control of listeria through intracellular innate immune recognition in drosophila (2008)
figf5

(a) Dot- or ring-shaped GFP-LC3 (LC3 dot) signals in S2 cells expressing both PGRP-LE and GFP-LC3 under the control of an actin promoter (PGRP-LE GFP-LC3) or GFP-LC3 alone (GFP-LC3) after infection with wild-type or Δhly L. monocytogenes or after 1.5 h of incubation with 5 μM rapamycin (rap). –, no infection. (b) Colocalization of GFP-LC3 dots and wild-type L. monocytogenes in S2 cells, quantified by confocal microscopy. (c) Confocal microscopy of S2 cells expressing PGRP-LE and GFP-LC3 (green) infected with wild-type L. monocytogenes and stained with DAPI (magenta). Arrow indicates colocalization of GFP-LC3 and L. monocytogenes. (d) Immunoblot analysis of lysates of S2 cells left untreated (–) or infected with wild-type L. monocytogenes or treated with 5 μM rapamycin. (e–i) Ultrastructural analysis of S2 cells expressing PGRP-LE and GFP-LC3 and infected with wild-type L. monocytogenes. (e,f) Fluorescence microscopy (e) and electron microscopy (f) of cells expressing GFP-LC3 (green) and stained with DAPI (magenta). (g) Enlargement of a bacteria-containing vacuole from f. (h,i) Enlargement of the fields outlined in g. Arrows indicate double-membrane structure that surrounds the bacteria; arrowheads indicate an endoplasmic reticulum–like membrane. (j) Confocal microscopy of hemocytes infected with wild-type L. monocytogenes. Green, GFP-LC3; magenta, DAPI. Arrow indicates colocalization of GFP-LC3 and L. monocytogenes. (k) Dot- or ring-shaped GFP-LC3 signals in ex vivo–cultured hemocytes expressing GFP-LC3 (infection and treatment, horizontal axis). (l) Immunoblot analysis of lysates of hemocytes from third instar larvae cultured ex vivo and infected with L. monocytogenes or treated with rapamycin. Arrowhead indicates processed form of GFP-LC3. (m) GFP-LC3 dots in S2 cells expressing PGRP-LE and GFP-LC3 and treated with RNAi (below graph) and infected for 0.5 h with L. monocytogenes then incubated for 1 h in gentamicin-containing medium; dots quantified by confocal microscopy. Scale bars, 5 μm (c,j), 1 μm (e,f) or 500 nm (g). *, P 0.001 (t-test). Data are representative of three (a,k,m), two (b,e–i), six (c), four (d) or five (j,l) experiments (error bars, s.d. of triplicate measurements (a,b,l) or at least triplicate measurements (m)).
Autophagic control of listeria through intracellular innate immune recognition in drosophila (2008)
figf6

(a) GFP-LC3 dots in S2 cells expressing GFP-LC3 alone or PGRP-LE and GFP-LC3, treated for 2 h with TCT (100 nM), highly purified DAP-type PGN from L. plantarum (DAP; 100 μg/ml) or lysine-type PGN from S. epidermidis (Lys). (b) Dot- or ring-shaped GFP-LC3 signals in hemocytes after infection with wild-type or Δhly L. monocytogenes or treatment with rapamycin (5 μM) or with TCT, DAP-type PGN or lysine-type PGN. *, P 0.001 (t-test). Data are representative of two independent experiments (error bars, s.d.).
Phospholipase Da6 and phosphatidic acid regulate gibberellin signaling in rice (2021)
Figure 4

Fig. 4. PA-GID1 interaction and amino acid residues involved in the binding.

Immunoblotting of His-GID1 and His-SLR1 expressed in E. coli.

Lipid-protein blotting assay of PA, PC, PE, PG, and PS with GID1 and SLR1. Lipids (0.5 μg) were spotted on nitrocellulose strips. PA, PC, PE, and PG were from egg yolk, and PS was from porcine brain. Purified proteins (GID1 and SLR1, 0.5 mg/ml) were used, followed by immunoblotting with anti-His-tag antibodies and color development.

Liposomes were made from di18:1-PC only or di18:1-PA/PC (PA:PC = 1:3 mole ratio). 1× and 10× refer to the concentration of PC or PA/PC liposomes used. NL, no liposome was added to the binding mixture.

Schematic diagram showing serial deletions of GID1. GID1 fragments were expressed in E. coli and used for defining the PA binding region. CD1/2 denote conserved domain HGG and GXSXG. Catalytic triad including three conserved amino acids of GID1, S, D, and H (vertical red bars).

Immunoblotting of His-GID1 proteins using constructs shown in (D). Proteins were separated by SDS-PAGE, followed by immunoblotting with anti-His-tag antibodies. The PA binding activity of different truncation mutations were analyzed by fat-immunoblotting. The red arrowheads indicate truncated proteins with different molecular weights.

Sequence alignment of the PA-binding fragment of GID1 with that of the PA binding motifs in chicken Raf1, abscisic acid insensitive 1 (ABI1), constitutive triple response1 (CTR1) and werewolf (WER) from Arabidopsis. Residues in bold are basic, potentially involved in PA binding and were mutated to Ala in GID1.

Immunoblotting of His-GID1 mutants and lipid-immunoblotting of PA binding by GID1 mutant proteins on a filter.

Liposomal binding of GID1 proteins to PA. Liposomes were made from di18:1-PA/PC (PA:PC = 1:3 mole ratio). Liposomal associated proteins were subjected to SDS-PAGE and immunoblotting using anti-His antibodies. The band intensity was analyzed by Image J and the intensity of input was set as 100%.
Phospholipase Da6 and phosphatidic acid regulate gibberellin signaling in rice (2021)
Figure 5

Fig. 5. Subcellular localization of PLDα6.

PLDα6-GFP distribution in rice protoplasts with different concentrations of GA3 or IAA. Protoplasts from 12-day old rice (ZH11 background) leaf sheath tissue were collected. Rice protoplasts were transfected with pM999-PLDα6 for 12 hours. After that, GA3 or IAA was added to protoplasts and 2 hours later, confocal images of protoplasts are shown. pM999-GFP refers to the empty vector with GFP only that was transformed as control and Ghd7-RFP was a nucleus marker. Scale bar = 10 μm.

Immunoblotting of PLDα6 in subcellular fractions. Total (T), soluble (S) and nuclear (N) proteins were isolated from PLDα6:GFP expressed in rice protoplasts treated with 10 μM GA3 or IAA. Equal amount of each sample was loaded for SDS-PAGE and blotting.
Phospholipase Da6 and phosphatidic acid regulate gibberellin signaling in rice (2021)
Figure 6

Fig. 6. Effect of PLDα6-KO on subcellular localization of GID1.

Protoplasts from 12-day old rice (ZH11 background) leaf sheath tissue were collected. and protoplasts were transfected with pM999-GID1 and mutation constructs and imaged 12 hours after transfection. All confocal images were scanned using similar laser gain and offset settings. Bars = 10 μm.

Subcellular location of GID1 in WT and pldα6 protoplasts with or without GA3 treatments. Cells for 12 hours after transformation were treated with or without 10 μM GA3 for two hours. Bars = 10 μm.

Immunoblotting of subcellular fractions of GID1 and mutations expressed in rice protoplasts. Total (T), soluble (S) and nuclear (N) protein fractions were isolated from protoplasts for 12 hours after transformation. Equal amounts of proteins from each sample were loaded.

Immunoblotting of subcellular fractions of GID1 expressed in WT and pldα6 protoplasts. After 2-hour treatment with 10 μM GA3, total (T), soluble (S) and nuclear (N) protein fractions were isolated from 10 samples of protoplasts. Equal amount of each sample was loaded for SDS-PAGE and immunoblotting.
Phospholipase Da6 and phosphatidic acid regulate gibberellin signaling in rice (2021)
Figure 7

Fig. 7. SLR protein stability in WT and pldα6 protoplasts.

Protoplasts of WT and pldα6 from 12-day old stage leaf sheath were collected and transfected with a pM999-SLR1 construct, incubated for 12 hours and then treated with 10 μM GA3 for 0, 3, and 9 hours. The first column, fluorescence from GFP; the second, red fluorescence from the nucleus marker Ghd7; the third, bright field, and the fourth, overlay of the three channels. All confocal images were scanned using similar laser gain and offset settings. Bars = 10 μm.

Immunoblotting of SLR1-GFP proteins from protoplast cells. Equal amounts of proteins from each sample were used for SDS-PAGE, followed by immunoblotting with anti-GFP antibodies.
miRNA profiling of human naive CD4 T cells links miR-34c-5p to cell activation and HIV replication (2016)
Figure 2

Figure 2. miR-34c-5p is up-regulated in naive CD4 T cells in response to TCR-mediated stimulation.

A. Average miR expression levels in naive CD4 T cells (purity >97%) before and after 72h TCR-stimulation. Lines indicate changes of +/- 1 log2 fold between samples (n=2, paired pools of three and nine individual samples per library). Library normalization performed using the mean normalization method available in Deseq package of Bioconductor (Anders & Huber, 2010). miRs with significant changes are highlighted (black) and with insets presenting mean read counts in unstimulated samples (mean), log2 fold change values in stimulated samples (log FC) and corresponding adjusted p value (p; Fisher test implemented in Deseq). The two most highly expressed miRs (mir-21-5p and miR-146b-5p) are also indicated.

B. Relative miR-34c-5p and miR-155-5p expression levels (2deltaCt normalized to RNU48) in unstimulated and stimulated naive CD4 T cells of individual samples pooled to generate the sequencing libraries. mean ± SEM and p-value for differences in miR-155-5p expression (paired two tailed t-test) are shown. Comparison could not be performed for miR-34c-5p as it is undetermined in unstimulated samples.

C. Time course quantification of miR-34c-5p expression (top panel, 2deltaCt values), cell proliferation (middle panel) and cell activation (bottom panel) markers by flow cytometry, in three different donors.
miRNA profiling of human naive CD4 T cells links miR-34c-5p to cell activation and HIV replication (2016)
Figure 3

Figure 3. Changes in miR expression in response to HIV infection of TCR-stimulated naive CD4 T cells.

A, B. Comparison of the mean miR expression level by in TCR-stimulated naive CD4 T cell small RNA-seq libraries from uninfected and HIV-1NL4-3 (A) or HIV-2ROD (B) infected samples (24h). Only miRNAs with a minimum of 10 normalized read counts. Library normalization was performed using the mean normalization method available in Deseq package of Bioconductor (Anders & Huber, 2010). Lines indicate changes of +/- 1 log2 fold between samples (n=3, paired pools of three individual samples per library). miR-34c-5p is highlighted in black.

C. Quantification of miR-34c-5p expression levels of the individual samples pooled to generate the sequencing libraries by qPCR (mean±SEM 2deltaCt values and p value from a paired two tailed t test; n=5).
miRNA profiling of human naive CD4 T cells links miR-34c-5p to cell activation and HIV replication (2016)
Figure 4

Figure 4. Impact of miR-34c-5p expression on HIV infection and replication.

A. Cell-associated viral DNA levels (left), Tat mRNA levels (center), and RT activity levels (right) in cell culture supernatants in infected J34c cells vs. infected JØ cells at 24 and 48 hours post-infection. * p0.05 (paired two tailed t test).

B. Representative flow cytometry plot of intracellular HIV-1 Gag protein staining (KC57) versus side scatter (SSC) in JØ and J34c cells, mock infected or infected with HIV-1NL4-3, after 48 hours of culture and corresponding fold change (J34c/JØ) plot in four biological replicate assays. Differences to control were found to be significant with a paired two tailed t test (p0.05).

C. Fold change levels for miR-34c-5p, cell-associated viral DNA level, Tat mRNA, and RT activity levels in J34c cells treated with miR-34c-5p antagomiR versus control antagomiR, 48 hours post-infection. Differences to control were found to be significant with a paired two tailed t test (p0.05).

All plots show fold change levels observed in a total of three biological replicate experiments performed (four for panel B). Lines depict the corresponding mean and SEM values.
miRNA profiling of human naive CD4 T cells links miR-34c-5p to cell activation and HIV replication (2016)
Figure 5

Figure 5. Impact of miR-34c-5p overexpression on Jurkat T cells.

A. Hierarchical clustering of the 82 differentially expressed (DE) genes identified in Jurkat cell lines overexpressing miR-34c-5p (J34c-2 and J34c-5) vs control (JØ-1, 2 and 3 clones). Color scale represents mean centered log2 fold expression level across samples. Pie charts represent the relative proportion of predicted miR-34c-5p binding sites by the Miranda or PITA algorithms (or both) in down-regulated (top) and up-regulated (bottom) gene sets.

B. qRT-PCR validation of selected genes. Plot represents the average percentage of target gene mRNA (normalized to GAPDH) in J34c vs JØ cells (all differences were found to be significant with a paired two tailed t test, P<0.05; n=5).