Journal: International Journal of Biological Sciences

Article Title: Targeting HSP90-HDAC6 Regulating Network Implicates Precision Treatment of Breast Cancer

doi: 10.7150/ijbs.18834

Figure Lengend Snippet: Combination of HDAC6 and HSP90 inhibition showed synergistic effect on ERα+ BC T47D cells

Article Snippet: Comparison of HDAC6 expression between ERα+ breast cancer cell lines and Basal breast cancer cell lines was made by applying Graphpad.

Techniques: Inhibition

Journal: International Journal of Biological Sciences

Article Title: Targeting HSP90-HDAC6 Regulating Network Implicates Precision Treatment of Breast Cancer

doi: 10.7150/ijbs.18834

Figure Lengend Snippet: Effects on microtubule cytoskeleton network stabilization of breast cancer cells upon inhibition of HSP90 or HDAC6 or both HSP90 and HDAC6. A. Treatment of 0.1μM 17-DMAG for 24 hours decreased HDAC6 protein level in both ERα+ breast cancer cells and TNBC cells. B. Inhibition of HSP90-HDAC6 with 0.1μM 17-DMAG and 1μM Tubacin increased tubulin acetylation synergistically. C. HSP90/HDAC6 inhibition with 0.1μM 17-DMAG and 1μM Tubacin decreased cell migration in both ERα+ breast cancer cells and TNBC cells. D/E. Analysis and comparison the effects on microtubule cytoskeleton stability in breast cancer cells upon inhibition of HSP90 or HDAC6 or both HSP90 and HDAC6. Each group was compared with control group, *P<0.05.

Article Snippet: Comparison of HDAC6 expression between ERα+ breast cancer cell lines and Basal breast cancer cell lines was made by applying Graphpad.

Techniques: Inhibition, Migration, Comparison, Control

Journal: International Journal of Biological Sciences

Article Title: Targeting HSP90-HDAC6 Regulating Network Implicates Precision Treatment of Breast Cancer

doi: 10.7150/ijbs.18834

Figure Lengend Snippet: Effects on ERK, AKT and Hippo pathway in breast cancer cells upon inhibition of HSP90. A. Investigation of AKT (p-AKT), ERK (p-ERK), and YAP (p-YAP) levels in TNBC cells and ERα positive breast cancer cells upon 0.1μM and 1μM HSP90 inhibitor 17-DMAG treatment. B. Inhibition of HDAC6 with 1μM Tubacin led to a decrease of YAP with an increase in tubulin acetylation, AKT and ERK phosphorylation. C. Effects on cell viability of breast cancer cells upon YAP silencing using siRNAs. Each group was compared to control group, *P<0.05.

Article Snippet: Comparison of HDAC6 expression between ERα+ breast cancer cell lines and Basal breast cancer cell lines was made by applying Graphpad.

Techniques: Inhibition, Phospho-proteomics, Control

Journal: International Journal of Biological Sciences

Article Title: Targeting HSP90-HDAC6 Regulating Network Implicates Precision Treatment of Breast Cancer

doi: 10.7150/ijbs.18834

Figure Lengend Snippet: Effects on HDAC6 expression in breast cancer cells upon inhibition or depletion ERα and in tamoxifen-resistant breast cancer cells A. Inhibition of ERα with 1μM tamoxifen or 0.1μM fulvestrant for 24 hours up-regulated HDAC6 at protein level in ERα positive breast cancer cells. B. Silencing ERα with siRNA elevated HDAC6 protein levelin ERα+ breast cancer cells. C. Inhibition of ERα with 1μM tamoxifen or 0.1μM fulvestrant for 24 hours elevated YAP expression at protein level. D. Higher HDAC6 and YAP expression with lower ERα and acetylated tubulin in T47D-TAR cells compared to those in their parental T47D cells.

Article Snippet: Comparison of HDAC6 expression between ERα+ breast cancer cell lines and Basal breast cancer cell lines was made by applying Graphpad.

Techniques: Expressing, Inhibition

Journal: International Journal of Biological Sciences

Article Title: Targeting HSP90-HDAC6 Regulating Network Implicates Precision Treatment of Breast Cancer

doi: 10.7150/ijbs.18834

Figure Lengend Snippet: Proposed model for the mechanism of HSP90-HDAC6 regulating network and implication in breast cancer treatment A-C. Proposed molecular mechanism driving breast cancer cells sensitive to HSP90 inhibition and HDAC6 inhibition. D. HSP90 inhibitor 17-DMAG inhibited tumor growth in TNBC cell xenograft mouse model. E. HDAC6 inhibitor Tubacin inhibited tumor growth in tamoxifen-resistant breast cancer cell xenograft mouse model. Each group was compared with control group, *P<0.05.

Article Snippet: Comparison of HDAC6 expression between ERα+ breast cancer cell lines and Basal breast cancer cell lines was made by applying Graphpad.

Techniques: Inhibition, Control

Journal: PLoS ONE

Article Title: Overexpression of Histone Deacetylase 6 Enhances Resistance to Porcine Reproductive and Respiratory Syndrome Virus in Pigs

doi: 10.1371/journal.pone.0169317

Figure Lengend Snippet: (A) Schematic diagram of the transgenic vector (pCMV-pHDAC6-Puro). The pair of black arrows indicates the primers used to identify the transgenic insert in TG pigs and in fibroblast colonies. The pair of green arrows indicates the primers used to identify exogenous HDAC6 mRNA in pigs. The pair of red arrows indicates the primers used to determine total HDAC6 mRNA levels in F1 pigs. (B) PCR assay to detect founder transgenic pigs. The PCR product (771 bp) was the GFP tag of the inserted transgene, which was amplified using the HDAC6-F/R primer pair. M, 100 bp DNA ladder; Lanes 1–9, founders No. 1–9; w, water; P, plasmid control; N, wild-type pig genomic DNA, used as the negative control. (C) qRT-PCR analysis of exogenous HDAC6 expression in different tissues (blood, liver, kidney, skin, lung, intestine and spleen) of the F0 transgenic pig using the Q-GFP-F/R primer pair. The blood sample data are presented as the mean±SD from 3 individuals (No. 1, 2 and 3). The data from other tissues are presented as the mean±SD from 3 repeated experiments. RNA from the intestines was used as the reference sample. WT, wild-type control. (D) Western blot analysis of F0 transgenic pigs. The samples were collected from tissues of TG (No.1) and WT pigs. The protein samples were probed with an anti-GFP antibody.

Article Snippet: Fibroblasts were transfected with linearized pig HDAC6 expression vectors (digested with Sca I) using an Amaxa Nucleofector Kit (Lonza) according to the manufacturer’s instructions.

Techniques: Transgenic Assay, Plasmid Preparation, Amplification, Control, Negative Control, Quantitative RT-PCR, Expressing, Western Blot

Journal: PLoS ONE

Article Title: Overexpression of Histone Deacetylase 6 Enhances Resistance to Porcine Reproductive and Respiratory Syndrome Virus in Pigs

doi: 10.1371/journal.pone.0169317

Figure Lengend Snippet: (A) qRT-PCR analysis of exogenous HDAC6 expression in the lungs and skin of F1 TG (n = 9) and sibling NTG pigs (lung, n = 10; skin, n = 6) using the Q-GFP-F/R primer pair. The data are presented as the mean ± SD. The relative expression of GFP was calculated using the 2 -Δct method. (B) qRT-PCR analysis of HDAC6 expression in lungs of F1 TG (n = 9) and sibling NTG pigs (n = 10) using the Q-HDAC6-F/R primer pair. (C) qRT-PCR analysis of HDAC6 expression in the skin of F1 TG (n = 9) and sibling NTG pigs (n = 6) using the Q-HDAC6-F/R primer pair. The data are presented as the mean±SD. The mRNA relative expression of total HDAC6 was calculated using the 2 -ΔΔct method (RNA from NTG pigs was used as the reference sample). GAPDH was used as an internal qRT-PCR control. Statistical significance was analyzed using a t-test. ***, P<0.001 **; P<0.01. (D) Western blot analysis of F1 pigs. The samples were collected from lung biopsies of six TG pigs and three sibling NTG pigs. The protein samples were probed with an anti-GFP antibody. GAPDH was used as an internal control for western blot analysis.

Article Snippet: Fibroblasts were transfected with linearized pig HDAC6 expression vectors (digested with Sca I) using an Amaxa Nucleofector Kit (Lonza) according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Expressing, Control, Western Blot

Journal: PLoS ONE

Article Title: Overexpression of Histone Deacetylase 6 Enhances Resistance to Porcine Reproductive and Respiratory Syndrome Virus in Pigs

doi: 10.1371/journal.pone.0169317

Figure Lengend Snippet: (A) qRT-PCR analysis of HDAC6 expression in PAMs isolated from pigs using the Q-HDAC6-F/R primer pair. HDAC6 expression is presented as a ratio relative to the level of GAPDH . The data are presented as the mean±SD from three independent experiments. (B) Western blot analysis of PAMs isolated from F1 pigs. The protein samples were probed with anti-Actub and GAPDH antibodies. (C) qRT-PCR analysis of viral ORF7 RNA levels in TG and NTG PAMs that were inoculated with the PRRSV strain CH-1a (MOI = 0.5) for 24, 48 and 72 h. The data represent the results of three independent experiments (mean±SD). (D) qRT-PCR analysis of viral ORF7 RNA levels in TG and NTG PAMs that were inoculated with the PRRSV strain JXA1 (MOI = 0.25) for 24 and 48 h. The data are presented relative to the expression of GAPDH mRNA and represent the results of three independent experiments (mean±SD). RNA from NTG PAMs at 24 hpi was used as the reference sample. Statistical significance was analyzed using a t-test. *, P<0.05; **, P<0.01; ***; P<0.001. (E) The supernatant containing PRRSV RNA was analyzed based on absolute quantitative RT-PCR values at the indicated time points. PAMs were infected with PRRSV CH-1a (MOI = 0.05), and the supernatant was collected and used for RNA extraction and absolute qPCR analysis of the virions at 24, 48 and 72 hpi. The data are representative of the results of three independent experiments (mean±SD). Statistical significance was analyzed using Student’s t-test. **, P<0.01; ***, P<0.001.

Article Snippet: Fibroblasts were transfected with linearized pig HDAC6 expression vectors (digested with Sca I) using an Amaxa Nucleofector Kit (Lonza) according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Expressing, Isolation, Western Blot, Infection, RNA Extraction

Journal: PLoS ONE

Article Title: Overexpression of Histone Deacetylase 6 Enhances Resistance to Porcine Reproductive and Respiratory Syndrome Virus in Pigs

doi: 10.1371/journal.pone.0169317

Figure Lengend Snippet: Kinetic analysis of JXA1-induced α-tubulin acetylation in MARC-145 cells (A) and in untreated cells (B). Kinetic analysis of JXA1-induced α-tubulin acetylation in PAMs (C) and in untreated cells (D). Kinetic analysis of JXA1-induced α-tubulin acetylation in HDAC6-transfected MARC-145 cells (E) and in untreated cells (F). The α-tubulin acetylation or α-tubulin content was quantified and is presented as a ratio relative to the total amount of GAPDH (Actub/GAPDH or tub/GAPDH).

Article Snippet: Fibroblasts were transfected with linearized pig HDAC6 expression vectors (digested with Sca I) using an Amaxa Nucleofector Kit (Lonza) according to the manufacturer’s instructions.

Techniques: Transfection