Journal: BMC Medical Genomics

Article Title: Integrated multi-omic analysis and experiment reveals the role of endoplasmic reticulum stress in lung adenocarcinoma

doi: 10.1186/s12920-023-01785-4

Figure Lengend Snippet: The primer sequence of 6 genes

Article Snippet: In brief, the tissue sections were incubated with a primary Anti-NUPR1 antibody (dilution 1:100; catalog number 15056-1-AP, Proteintech, China) and subsequently processed using the appropriate detection system.

Techniques: Sequencing

Journal: BMC Medical Genomics

Article Title: Integrated multi-omic analysis and experiment reveals the role of endoplasmic reticulum stress in lung adenocarcinoma

doi: 10.1186/s12920-023-01785-4

Figure Lengend Snippet: Single cell sequencing analysis of ERSRGs-signature. ( A ) tSNE clustering colored by groups. ( B ) The annotation of clusters based on marker analysis. mRNA distribution of BAK1 ( C ), EIF2AK3 ( D ), MBTPS2 ( E ), NUPR1 ( F ), RHBDD2 ( G ) and VCP ( H ) after tSNE dimensionality reduction. ( I ) Differential expression of ERSRGs in the different cell clusters

Article Snippet: In brief, the tissue sections were incubated with a primary Anti-NUPR1 antibody (dilution 1:100; catalog number 15056-1-AP, Proteintech, China) and subsequently processed using the appropriate detection system.

Techniques: Sequencing, Marker, Quantitative Proteomics

Journal: BMC Medical Genomics

Article Title: Integrated multi-omic analysis and experiment reveals the role of endoplasmic reticulum stress in lung adenocarcinoma

doi: 10.1186/s12920-023-01785-4

Figure Lengend Snippet: Validation of the expression levels of ERSRGs in LUAD. The mRNA expression of BAK1 ( A ), EIF2AK3 ( B ), MBTPS2 ( C ), NUPR1 ( D ), RHBDD2 ( E ) and VCP ( F ) in LUAD patients from Nantong tumor hospital. N = 8, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ( G ) The protein expression of BAK1, EIFAK3, MBTPS2, RHBDD2 and VCP in HPA

Article Snippet: In brief, the tissue sections were incubated with a primary Anti-NUPR1 antibody (dilution 1:100; catalog number 15056-1-AP, Proteintech, China) and subsequently processed using the appropriate detection system.

Techniques: Biomarker Discovery, Expressing

Journal: BMC Medical Genomics

Article Title: Integrated multi-omic analysis and experiment reveals the role of endoplasmic reticulum stress in lung adenocarcinoma

doi: 10.1186/s12920-023-01785-4

Figure Lengend Snippet: Expression analysis of NUPR1 at transcription and translation Levels. Representative images ( A ) and quantification ( B ) of NUPR1 in intratumoral and peritumoral fractions through immunohistochemistry staining (N = 6). MRNA ( C ) and protein expression ( D & E ) of NUPR1 in cell lines (N = 3). ( F ) Cell viability assessed through CCK8 assays between saline and trifluoperazine subgroups (N = 6). ( G ) Representative images and results of cell counting from the Transwell invasion assay (N = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: In brief, the tissue sections were incubated with a primary Anti-NUPR1 antibody (dilution 1:100; catalog number 15056-1-AP, Proteintech, China) and subsequently processed using the appropriate detection system.

Techniques: Expressing, Immunohistochemistry, Staining, Saline, Cell Counting, Transwell Invasion Assay

Journal: Journal of Hypertension

Article Title: Rho kinase inhibition ameliorates vascular remodeling and blood pressure elevations in a rat model of apatinib-induced hypertension

doi: 10.1097/hjh.0000000000003060

Figure Lengend Snippet: FIGURE 2 Effect of apatinib treatment on level of RhoA and Rho-associated coiled-coil domain-containing protein kinase mRNAs. Apatinib upregulated RhoA and ROCK II mRNAs relative to the vehicle (P < 0.001) but downregulated GRAF3 (P < 0.001) in the mid-aorta. Apatinib had no effect on level of ROCK I mRNAs between vehicle and treatment groups (n ¼ 8 in each group). P less than 0.05. ROCK, Rho-associated coiled-coil domain-containing protein kinase.

Article Snippet: The membranes were blocked with 5% skim milk, and immunoblotted with the following primary antibodies: RhoA (1 : 1000, 10749–1-AP, Protein-tech, Wuhan, China), ROCK I (1 : 1000, ab134181, Abcam, Cambridge, UK), ROCK II (1 : 20000, ab125025, Abcam), MLCP (1 : 1000, ab32519, Abcam), endothelin A receptor (ETA, 1 : 900, ab85163, Abcam), and collagen I (Col I, 1 : 500, ab6308, Abcam).

Techniques:

Journal: Journal of Hypertension

Article Title: Rho kinase inhibition ameliorates vascular remodeling and blood pressure elevations in a rat model of apatinib-induced hypertension

doi: 10.1097/hjh.0000000000003060

Figure Lengend Snippet: FIGURE 3 Effect of apatinib on expression of proteins in the RhoA/Rho-associated coiled-coil domain-containing protein kinase signaling pathway. Apatinib upregu- lated expression of RhoA and ROCK II proteins but had no significant effect on ROCK I expression in all groups (n ¼ 8 in each group). P less than 0.05. ROCK, Rho-associated coiled-coil domain-containing protein kinase.

Article Snippet: The membranes were blocked with 5% skim milk, and immunoblotted with the following primary antibodies: RhoA (1 : 1000, 10749–1-AP, Protein-tech, Wuhan, China), ROCK I (1 : 1000, ab134181, Abcam, Cambridge, UK), ROCK II (1 : 20000, ab125025, Abcam), MLCP (1 : 1000, ab32519, Abcam), endothelin A receptor (ETA, 1 : 900, ab85163, Abcam), and collagen I (Col I, 1 : 500, ab6308, Abcam).

Techniques: Expressing

Journal: Cancer research

Article Title: TUSC4 functions as a tumor suppressor by regulating BRCA1's stability via the E3 ubiquitination pathway

doi: 10.1158/0008-5472.CAN-14-2315

Figure Lengend Snippet: A. Lower TUSC4 expression levels was found in both luminal and basal types of breast cancer cell lines, while non-transformed breast cell lines (HMEC, MCF-10A and MCF-12A) exhibited higher TUSC4 level.

Article Snippet: An anti-TUSC4 antibody was purchased from Proteintech (10157-1-AP), an anti-BRCA1 antibody was purchased from Santa Cruz Biotechnology (sc-6954), and anti-Flag M2 (F3165) and anti-β-actin (A2066) antibodies were purchased from Sigma.

Techniques: Expressing, Transformation Assay

Journal: Cancer research

Article Title: TUSC4 functions as a tumor suppressor by regulating BRCA1's stability via the E3 ubiquitination pathway

doi: 10.1158/0008-5472.CAN-14-2315

Figure Lengend Snippet: A. TUSC4 knockdown reduced BRCA1 foci formation after IR, while control cells didn't have such effect (NT). γ-H2AX indicated the efficiency of irradiation.

Article Snippet: An anti-TUSC4 antibody was purchased from Proteintech (10157-1-AP), an anti-BRCA1 antibody was purchased from Santa Cruz Biotechnology (sc-6954), and anti-Flag M2 (F3165) and anti-β-actin (A2066) antibodies were purchased from Sigma.

Techniques: Knockdown, Control, Irradiation

Journal: Cancer research

Article Title: TUSC4 functions as a tumor suppressor by regulating BRCA1's stability via the E3 ubiquitination pathway

doi: 10.1158/0008-5472.CAN-14-2315

Figure Lengend Snippet: A. Knockdown of TUSC4 didn't significantly change the cell cycle distribution of U2OS cells compared to control cells. G1, G2/M and S phases cells were indicated by percentage of total cell numbers.

Article Snippet: An anti-TUSC4 antibody was purchased from Proteintech (10157-1-AP), an anti-BRCA1 antibody was purchased from Santa Cruz Biotechnology (sc-6954), and anti-Flag M2 (F3165) and anti-β-actin (A2066) antibodies were purchased from Sigma.

Techniques: Knockdown, Control

Journal: Cancer research

Article Title: TUSC4 functions as a tumor suppressor by regulating BRCA1's stability via the E3 ubiquitination pathway

doi: 10.1158/0008-5472.CAN-14-2315

Figure Lengend Snippet: A. TUSC4 knockdown increased ubiquitination level of BRCA1 compared to control cells (After MG132 enrichment for ubiquitination).

Article Snippet: An anti-TUSC4 antibody was purchased from Proteintech (10157-1-AP), an anti-BRCA1 antibody was purchased from Santa Cruz Biotechnology (sc-6954), and anti-Flag M2 (F3165) and anti-β-actin (A2066) antibodies were purchased from Sigma.

Techniques: Knockdown, Ubiquitin Proteomics, Control

Journal: Cancer research

Article Title: TUSC4 functions as a tumor suppressor by regulating BRCA1's stability via the E3 ubiquitination pathway

doi: 10.1158/0008-5472.CAN-14-2315

Figure Lengend Snippet: A. Colony formation assay indicated that normal U2OS cells are not sensitive to PARP inhibitor Olaparib (1μm) but TUSC4-knockdown cells exhibited increased sensitivity to Olaparib, the colonies were significantly reduced after the treatment (p<0.05).

Article Snippet: An anti-TUSC4 antibody was purchased from Proteintech (10157-1-AP), an anti-BRCA1 antibody was purchased from Santa Cruz Biotechnology (sc-6954), and anti-Flag M2 (F3165) and anti-β-actin (A2066) antibodies were purchased from Sigma.

Techniques: Colony Assay, Knockdown

Journal: Cancer research

Article Title: TUSC4 functions as a tumor suppressor by regulating BRCA1's stability via the E3 ubiquitination pathway

doi: 10.1158/0008-5472.CAN-14-2315

Figure Lengend Snippet: Tumorigenicity of Orthotopically Implanted Control and TUSC4-overexpression MDA-MB-231 cells 5×10 6 cells from MDA-AB-231 control and two independent TUSC4-overexpressing MDA-AB-231 cell lines (TUSC4 #7 and TUSC4 #13) were injected per mouse into mammary tumor sizes were analyzed.

Article Snippet: An anti-TUSC4 antibody was purchased from Proteintech (10157-1-AP), an anti-BRCA1 antibody was purchased from Santa Cruz Biotechnology (sc-6954), and anti-Flag M2 (F3165) and anti-β-actin (A2066) antibodies were purchased from Sigma.

Techniques: Control, Injection

Journal: Cancer research

Article Title: TUSC4 functions as a tumor suppressor by regulating BRCA1's stability via the E3 ubiquitination pathway

doi: 10.1158/0008-5472.CAN-14-2315

Figure Lengend Snippet: Tumorigenicity of Orthotopically Implanted Control and TUSC4-Knockdown MCF10A cells 1×10 7 cells from MCF-10A control and two independent TUSC4-knockdown MCF-10A cell lines (TUSC4 #1 and TUSC4 #4) were injected per mouse into mammary fat pads glands of 6-week-old female nude mice. Each cell line was injected in five different mice, and tumor sizes were analyzed.

Article Snippet: An anti-TUSC4 antibody was purchased from Proteintech (10157-1-AP), an anti-BRCA1 antibody was purchased from Santa Cruz Biotechnology (sc-6954), and anti-Flag M2 (F3165) and anti-β-actin (A2066) antibodies were purchased from Sigma.

Techniques: Control, Injection

Journal: Frontiers in physiology

Article Title: TMEM16A Plays an Insignificant Role in Myocardium Remodeling but May Promote Angiogenesis of Heart During Pressure-overload.

doi: 10.3389/fphys.2022.897619

Figure Lengend Snippet: FIGURE 1 | Molecular expression levels of TMEM16A in LV. (A) TMEM16A mRNA expression level in LV (n = 3 mice in each group per time point, RM two-way ANOVA). (B) Traditional western blot results of TMEM16A protein expression level in LV (n = 3 mice in each group per time point, RM two-way ANOVA). (C) Simple western blot results of TMEM16A protein expression level in LV (n = 3 mice in each group per time point, RM two-way ANOVA).

Article Snippet: Briefly, total protein was separated on 10% SDS-PAGE, transferred onto a PVDF membrane and blotted with the following primary antibodies separately: Anti-TMEM16A antibody (ACL-011, Alomone labs) for protein extracted from mouse heart, Anti-TMEM16A antibody (BA3464-2, Boster) for endogenous human TMEM16A protein in HUVECs, AntiTMEM16A antibody (ab53212, Abcam) for overexpressed mouse TMEM16A protein induced by lentivirus in HUVECs, and Anti-β-actin antibody (CST).

Techniques: Expressing, Western Blot, Simple Western

Journal: Frontiers in physiology

Article Title: TMEM16A Plays an Insignificant Role in Myocardium Remodeling but May Promote Angiogenesis of Heart During Pressure-overload.

doi: 10.3389/fphys.2022.897619

Figure Lengend Snippet: FIGURE 2 | Ito in LVMs. (A) Recordings show the effects of T16Ainh-A01 (30 μM) or 4-AP (5 mM) on Ito in LVMs. (B) Recordings show the effects of Anti-TMEM16A antibody (1:200) or 4-AP (5 mM) on Ito in LVMs. (C) I-V relations of T16Ainh-A01 sensitive current and Anti-TMEM16A antibody sensitive current (n = 4 cells in each group, two-way ANOVA). (D) I-V relation of 4-AP sensitive current (n = 4 cells in each group, two-way ANOVA).

Article Snippet: Briefly, total protein was separated on 10% SDS-PAGE, transferred onto a PVDF membrane and blotted with the following primary antibodies separately: Anti-TMEM16A antibody (ACL-011, Alomone labs) for protein extracted from mouse heart, Anti-TMEM16A antibody (BA3464-2, Boster) for endogenous human TMEM16A protein in HUVECs, AntiTMEM16A antibody (ab53212, Abcam) for overexpressed mouse TMEM16A protein induced by lentivirus in HUVECs, and Anti-β-actin antibody (CST).

Techniques:

Journal: Frontiers in physiology

Article Title: TMEM16A Plays an Insignificant Role in Myocardium Remodeling but May Promote Angiogenesis of Heart During Pressure-overload.

doi: 10.3389/fphys.2022.897619

Figure Lengend Snippet: FIGURE 5 | Molecular expression levels of TMEM16A in HUVECs. (A) TMEM16A mRNA expression level in HUVECs (n = 3–4). (B) Western blot results of TMEM16A protein expression level in HUVECs (n = 3–5). *p < 0.05, **p < 0.01 and ***p < 0.001, one-way ANOVA and unpaired t-test.

Article Snippet: Briefly, total protein was separated on 10% SDS-PAGE, transferred onto a PVDF membrane and blotted with the following primary antibodies separately: Anti-TMEM16A antibody (ACL-011, Alomone labs) for protein extracted from mouse heart, Anti-TMEM16A antibody (BA3464-2, Boster) for endogenous human TMEM16A protein in HUVECs, AntiTMEM16A antibody (ab53212, Abcam) for overexpressed mouse TMEM16A protein induced by lentivirus in HUVECs, and Anti-β-actin antibody (CST).

Techniques: Expressing, Western Blot

Journal: Frontiers in physiology

Article Title: TMEM16A Plays an Insignificant Role in Myocardium Remodeling but May Promote Angiogenesis of Heart During Pressure-overload.

doi: 10.3389/fphys.2022.897619

Figure Lengend Snippet: FIGURE 6 | Effect of TMEM16A on migration in HUVECs. (A) Wound healing assay results of HUVECs transfected with siRNANC or siRNAMix, and treated with vehicle (PBS), or VEGF (30 ng/mL), or AngII (1 μM) separately (n = 5—8 in each group per time point). (B) Wound healing assay results of HUVECs transfected with TMNC

Article Snippet: Briefly, total protein was separated on 10% SDS-PAGE, transferred onto a PVDF membrane and blotted with the following primary antibodies separately: Anti-TMEM16A antibody (ACL-011, Alomone labs) for protein extracted from mouse heart, Anti-TMEM16A antibody (BA3464-2, Boster) for endogenous human TMEM16A protein in HUVECs, AntiTMEM16A antibody (ab53212, Abcam) for overexpressed mouse TMEM16A protein induced by lentivirus in HUVECs, and Anti-β-actin antibody (CST).

Techniques: Migration, Wound Healing Assay, Transfection

Journal: Frontiers in physiology

Article Title: TMEM16A Plays an Insignificant Role in Myocardium Remodeling but May Promote Angiogenesis of Heart During Pressure-overload.

doi: 10.3389/fphys.2022.897619

Figure Lengend Snippet: FIGURE 7 | Effect of TMEM16A on angiogenesis in HUVECs. (A,C) Tube formation assay results of HUVECs transfected with siRNANC or siRNAMix, or TMNC, or TMOE, and treated with vehicle (PBS), or VEGF (30 ng/mL), or AngII (1 μM) separately (n = 3–9). (B,D) Endothelial cell spheroids sprouting assay results of HUVECs transfected with siRNANC or siRNAMix, or TMNC, or TMOE, and treated with vehicle (PBS), or VEGF (30 ng/mL), or AngII (1 μM) separately (n = 3–5). *p < 0.05 compared with siRNANC or TMNC under the same treatment, #p < 0.05 and ##p < 0.01 compared with vehicle (PBS) in the same cell type, two-way ANOVA.

Article Snippet: Briefly, total protein was separated on 10% SDS-PAGE, transferred onto a PVDF membrane and blotted with the following primary antibodies separately: Anti-TMEM16A antibody (ACL-011, Alomone labs) for protein extracted from mouse heart, Anti-TMEM16A antibody (BA3464-2, Boster) for endogenous human TMEM16A protein in HUVECs, AntiTMEM16A antibody (ab53212, Abcam) for overexpressed mouse TMEM16A protein induced by lentivirus in HUVECs, and Anti-β-actin antibody (CST).

Techniques: Tube Formation Assay, Transfection

Journal: Frontiers in cell and developmental biology

Article Title: LncRNA NORAD Promotes Vascular Endothelial Cell Injury and Atherosclerosis Through Suppressing VEGF Gene Transcription via Enhancing H3K9 Deacetylation by Recruiting HDAC6.

doi: 10.3389/fcell.2021.701628

Figure Lengend Snippet: FIGURE 3 | Nucleus lncRNA NORAD associates with HDAC6 via FUS in ox-LDL-treated HUVECs. (A) Representative images of WB following RAP performed by probe-targeting lncRNA NORAD in HUVECs. (B) Potential binding sites for FUS in lncRNA NORAD analyzed by ENCORI. (C) Representative images of Co-IP using an anti-FUS antibody in HUVECs. Rabbit IgG was used as negative control. (D) Representative images of WB following RAP performed by probe-targeting lncRNA NORAD in HUVECs transfected with control shRNA or shRNA-targeting lncRNA NORAD under ox-LDL treatment. In the bar graph, the protein level of HDAC6 was quantified and normalized by input. shRNA; shNORAD, shRNA-targeting lncRNA NORAD; NC, negative control; siCtrl, control siRNA; siFUS, FUS siRNA (***p < 0.001 vs. other groups). N = 3.

Article Snippet: Subsequently, about 300 μg of protein was incubated with 1 μg of FUS antibody (Proteintech, United States) and 25 μl of protein A/G magnetic beads for immunoprecipitation at 4◦C overnight.

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Negative Control, Transfection, Control, shRNA