primary antibody against creb5 8a5  (Abnova)

Journal: Neoplasia (New York, N.Y.)

Article Title: A trio of tumor suppressor miRNA downregulates CREB5 dependent transcription to modulate neoadjuvant hormonal therapy sensitivity

doi: 10.1016/j.neo.2022.100875

Figure Lengend Snippet: CREB5 is the direct target of miR-l42-3p, miR-150-5p and miR-342-3p. (A-D) The mRNA level of AR and PSA measured in LNCaP-ENZR and 22RV1 cells after transfection with indicated miRNAs mimics/inhibitor (RT-qPCR). (E) Heatmap displaying the relation of enrichment score of AR related signatures with miR-l42-3p, miR-150-5p and miR-342-3p expression according to TCGA data. The color represents the correlation coefficient and P-value are reported. The AR related signatures were downloaded from MsigDB of GSEA ( http://www.gsea-msigdb.org/gsea/msigdb/index.jsp ). Enrichment score were quantified by ssGSEA in R package GSVA. (F) A Venn diagram depicted sixteen genes that commonly targeted by indicated miRNAs, which based on four prediction algorithms (MiRWalk,miranda,RNA22 and Targetscan). (G) Heatmap displaying the expression of miRNAs target genes in Tewari cohort. (H-I) The protein level of CREB5 assessed in LNCaP cells by Western blot after transfection with indicated miRNAs mimics/inhibitor and relative control RNA. (J-K) The recognition sites for indicated miRNAs in CREB5 3′UTR region. (L-N) Luciferase reporter assay performed in HEK-293T cells. Cells were transiently co-transfected with the wild or mutated CREB5 3′UTR region together with corresponding miRNA mimics, respectively. After incubation for 48h, luciferase activities were measured. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Briefly, sections were incubated overnight with primary antibody against CREB5 (Abnova, 8A5, USA.

Techniques: Transfection, Quantitative RT-PCR, Expressing, Western Blot, Control, Luciferase, Reporter Assay, Incubation

Journal: Neoplasia (New York, N.Y.)

Article Title: A trio of tumor suppressor miRNA downregulates CREB5 dependent transcription to modulate neoadjuvant hormonal therapy sensitivity

doi: 10.1016/j.neo.2022.100875

Figure Lengend Snippet: CREB5 promotes proliferation and antiandrogen resistance in vitro . (A-D) Cell proliferation measured in LNCaP and VCaP cells. MTS assays (A-B) and EdU (C-D) assays were performed in LNCaP cells with CREB5 overexpression and in VCaP cells with CREB5 knockdown. Quantitative results of EdU assays are shown in the right panel. (E) Cell viability measured in the LNCaP cells under bicalutamide treatment. LNCaP cells were transfect with CREB5 plasmids/negative control and treated with titrated doses of bicalutamide for 48h. Cell viability was quantified by MTS assays. Data shown are means ± SD. (F) Cell proliferation determined with bicalutamide treatment. LNCaP cells were transiently transfected with CREB5 overexpression. Following treatment with 44 μM bicalutamide, the cells were subjected to MTS assays. (G-J) Relative proliferation rate of LNCaP-ENZR (G-H) and 22RV1 cells (I-J). Cells were transfected as indicated, then subjected to MTS assays treated with bicalutamide or enzalutamide. Cells were cultured in 10% CS-FBS. The absorbance at 490nm was measured after 48h treatment with bicalutamide or enzalutamide. (K-L) The mRNA levels of AR(K) and PSA(L) assessed by RT-qPCR after transfection as indicated in 22RV1 cells. * P < 0.05, ** P < 0.01, *** P < 0.001. All data represent means ± SD of at least three independent replicates.

Article Snippet: Briefly, sections were incubated overnight with primary antibody against CREB5 (Abnova, 8A5, USA.

Techniques: In Vitro, Over Expression, Knockdown, Negative Control, Transfection, Cell Culture, Quantitative RT-PCR

Journal: Neoplasia (New York, N.Y.)

Article Title: A trio of tumor suppressor miRNA downregulates CREB5 dependent transcription to modulate neoadjuvant hormonal therapy sensitivity

doi: 10.1016/j.neo.2022.100875

Figure Lengend Snippet: IL6 signaling is involved in CREB5-mediated antiandrogen resistance. (A-B) Gene sets significantly enriched in high-CREB5 tumors accordingly to the original annotation in the MsigDB Database Hallmark ( http://software.broadinstitute.org/gsea/msigdb/index.jsp ). Normalized enrichment score (NES) and P-value (P) are reported. (C-D) Correlation between the mRNA levels of CREB5 and IL6 in the GSE141551 (C) and TCGA data (D). Spearman r score and P value are shown. (E) The mRNA levels of IL6 assessed by RT-qPCR after transfection with CREB5 plasmids in LNCaP cells. (F-I) The mRNA levels of IL6 assessed by RT-qPCR after transfection with miRNAs mimics and inhibitors in 22RV1 (F-G) and LNCaP-ENZR (H-I) cells. * P < 0.05, ** P < 0.01.

Article Snippet: Briefly, sections were incubated overnight with primary antibody against CREB5 (Abnova, 8A5, USA.

Techniques: Software, Quantitative RT-PCR, Transfection

Journal: Neoplasia (New York, N.Y.)

Article Title: A trio of tumor suppressor miRNA downregulates CREB5 dependent transcription to modulate neoadjuvant hormonal therapy sensitivity

doi: 10.1016/j.neo.2022.100875

Figure Lengend Snippet: Clinicopathological analysis for CREB5 by immunohistochemistry in 85 PCa patients.

Article Snippet: Briefly, sections were incubated overnight with primary antibody against CREB5 (Abnova, 8A5, USA.

Techniques: Immunohistochemistry, Expressing, Significance Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: A trio of tumor suppressor miRNA downregulates CREB5 dependent transcription to modulate neoadjuvant hormonal therapy sensitivity

doi: 10.1016/j.neo.2022.100875

Figure Lengend Snippet: CREB5 is highly expressed in NHT-R PCa tissues. (A-D) CREB5 expression measured by IHC assay in Qilu cohort. Representative IHC images of CREB5 expression in PCa biopsy tissues with NHT-S (A1) or NHT-R (A2) in Qilu cohort. Quantitative analysis of cases with CREB5 positive was shown in A3. ROC curve was constructed to validate the sensitivity and specificity of CREB5 in predicting NHT-R (B). Representative IHC images of CREB5 expression in cases with Gleason score <8 (C1), and Gleason score ≥8 (C2-C3). ROC curve was constructed to validate the sensitivity and specificity of CREB5 in predicting GS (D). (E-G) The relevance between CREB5 and the three miRNAs expression in Qilu cohort. (H-K) Analysis of CREB5 expression in TCGA dataset. (H) CREB5 levels in PCa subgroups with different Gleason scores; (I) CREB5 levels in PCa subgroups with different clinical stages; (J) CREB5 levels in PCa subgroups with different pathological stages; (K) CREB5 levels in PCa subgroups with lymph node metastasis. (L-M) Analysis of CREB5 expression in MSKCC dataset. (L)The expression of CREB5 in primary and metastatic PCa tissues compared with the matched normal prostate samples in MSKCC dataset. (M) The correlation between CREB5 expression and biochemical recurrence-free survival in MSKCC cohort (Kaplan–Meier survival, Log-rank test). (N) The correlation between CREB5 expression and biochemical recurrence-free survival in GSE116918 data (Kaplan–Meier survival, Log-rank test). TCGA The Cancer Genome Atlas. GS, Gleason score. cT, clinical stage. pT, pathological stage. LNM, lymph node metastasis. BCR, biochemical recurrence. Error bars represent means ± SD of three independent experiments. Original magnification of IHC images, × 200. * P < 0.05.

Article Snippet: Briefly, sections were incubated overnight with primary antibody against CREB5 (Abnova, 8A5, USA.

Techniques: Expressing, Construct