Journal: Stem Cells International

Article Title: lncRNA TUG1 Facilitates Colorectal Cancer Stem Cell Characteristics and Chemoresistance by Enhancing GATA6 Protein Stability

doi: 10.1155/2021/1075481

Figure Lengend Snippet: Analysis of the interaction between lncRNA TUG1 and GATA6. (a) RNA Interactome Database and COAD colon cancer database were applied to predict the transcription factors that had potential interactions with lncRNA TUG1 and were highly expressed in CRC. (b) RNA pull-down assay was conducted to verify the interaction between lncRNA TUG1 and ETV4, ASCL2, GATA6, and SOX9. (c) Verification of the interaction between lncRNA TUG1 and GATA6 by RNA immunoprecipitation assay. (d, e) si-TUG1 was transfected into CD133 − /CD44 − or CD133 + /CD44 + isolated from SW480 cells. The mRNA and protein levels of GATA6 were analyzed using qRT-PCR and Western blot. (f, g) si-TUG1 was transfected into CD133 − /CD44 − or CD133 + /CD44 + isolated from SW480 cells, and the cells were treated with 100 μ g/ml cycloheximide (CHX) for 0, 4, and 8 h. Detection of the GATA6 protein level by Western blot. (h–k) si-TUG1 was transfected into CD133 − /CD44 − or CD133 + /CD44 + isolated from SW480 cells. The mRNA and protein levels of BMP4 and BMP2 were assessed using qRT-PCR and Western blot. ∗∗ P < 0.01 vs. si-NC or CD133 − /CD44 − . ∗∗∗ P < 0.001 vs. IgG or CD133 − /CD44 − .

Article Snippet: To assess the lncRNA TUG1 impact on the GATA6 protein degradation, si-TUG1 was transfected into CD133 − /CD44 − or CD133 + /CD44 + isolated from SW480 cells, and the cells were treated with 100 μ g/ml cycloheximide (CHX, #HY-12320, MedChemExpress) for 0, 4, and 8 h.

Techniques: Pull Down Assay, RNA Immunoprecipitation, Transfection, Isolation, Quantitative RT-PCR, Western Blot

Journal: Stem Cells International

Article Title: lncRNA TUG1 Facilitates Colorectal Cancer Stem Cell Characteristics and Chemoresistance by Enhancing GATA6 Protein Stability

doi: 10.1155/2021/1075481

Figure Lengend Snippet: Verification of the regulation of the lncRNA TUG1/GATA6 axis on the characteristics and chemoresistance of CRC stem cells. si-TUG1 and/or pcDNA-GATA6 were transfected into CD133 + /CD44 + isolated from SW480 cells. (a) qRT-PCR and Western blot assays were carried out to identify the transfection efficiency of pcDNA-GATA6. (b) Analysis of the protein levels of ALDH1 and Nanog in CD133 + /CD44 + isolated from SW480 cells using Western blot. (c) The formation of clonal spheres was assessed by sphere formation assay. si-TUG1 and/or pcDNA-GATA6 were transfected into CD133 + /CD44 + isolated from SW480 cells, and the cells were then treated with 4 μ M oxaliplatin for 48 h. (d) The viability of CD133 + /CD44 + isolated from SW480 cells was analyzed using a CCK-8 assay. (e) Analysis of the apoptotic ability of CD133 + /CD44 + isolated from SW480 cells by flow cytometry. ∗∗ P < 0.01 vs. si-NC. ∗∗∗ P < 0.001 vs. si-TUG1+pc-NC. # P < 0.05 and ### P < 0.001 vs. si-TUG1+pc-NC.

Article Snippet: To assess the lncRNA TUG1 impact on the GATA6 protein degradation, si-TUG1 was transfected into CD133 − /CD44 − or CD133 + /CD44 + isolated from SW480 cells, and the cells were treated with 100 μ g/ml cycloheximide (CHX, #HY-12320, MedChemExpress) for 0, 4, and 8 h.

Techniques: Transfection, Isolation, Quantitative RT-PCR, Western Blot, Tube Formation Assay, CCK-8 Assay, Flow Cytometry

Journal: Stem Cells International

Article Title: lncRNA TUG1 Facilitates Colorectal Cancer Stem Cell Characteristics and Chemoresistance by Enhancing GATA6 Protein Stability

doi: 10.1155/2021/1075481

Figure Lengend Snippet: Regulation of lncRNA TUG1 on the CRC stem cell characteristics and chemoresistance in vivo . CD133 + /CD44 + isolated from HCT-116 cells transfected with Lv-shTUG1 were injected into mice to establish a xenograft model and the oxaliplatin (5 mg/kg) was injected into mice every three days, and 6 mice were randomly assigned to each group. (a) The tumor volume of mice was monitored ( n = 6). (b) Detection of the lncRNA TUG1 expression in tumor tissues by qRT-PCR. (c) Detection of the GATA6 mRNA level using qRT-PCR. (d) Analysis of the GATA6, BMP4, and BMP2 protein levels in tumor tissues by Western blot. (e) Detection of the protein levels of ALDH1 and Nanog using Western blot. (f) The cell apoptosis in tumor tissues was assessed by TUNEL assay (scale bar: 20 μ m). ∗∗ P < 0.01 and ∗∗∗ P < 0.001 vs. Lv-shNC; ### P < 0.01 vs. Lv-shNC+OXA. OXA: oxaliplatin.

Article Snippet: To assess the lncRNA TUG1 impact on the GATA6 protein degradation, si-TUG1 was transfected into CD133 − /CD44 − or CD133 + /CD44 + isolated from SW480 cells, and the cells were treated with 100 μ g/ml cycloheximide (CHX, #HY-12320, MedChemExpress) for 0, 4, and 8 h.

Techniques: In Vivo, Isolation, Transfection, Injection, Expressing, Quantitative RT-PCR, Western Blot, TUNEL Assay

Journal: Bioscience Reports

Article Title: Hydrogen sulfide attenuates sepsis-induced cardiac dysfunction in infant rats by inhibiting the expression of cold-inducible RNA-binding protein

doi: 10.1042/BSR20241398

Figure Lengend Snippet: ( A–F ) Representative Western blots and quantitative analysis for GRP78, ATF6, p-PERK, PERK, p-IRE1-a, IRE1-a. GAPDH was used as the internal control. ( G ) Representative H&E staining left ventricular sections (scale bar = 50μm μm). ( H ) Severity scores of heart tissue sections. Data are mean ± SD from four independent experiments ( n = 6). One-way analysis of variance followed by LSD test with equal variance, and the Dunnett’s T3 method was performed if there was missing variance, P ＜0.05 was considered significant vs. sham + V. H&E, hematoxylin and eosin.

Article Snippet: The membranes were blocked with 3% BSA for 1.5 h at room temperature, and antigens were detected using the following antibodies at 4°C overnight: GRP78 (1:10000, Santa Cruz Biotechnology Inc., TX, U.S.A., sc-166490), ATF6 (1:500, MCE, SH, China, HY- P80379 ), p-PERK (1:1000, GeneTex, SZ, China, GTX00673), PERK (1:1000, Santa Cruz Biotechnology, TX, U.S.A., sc-377400), p-IRE1-a (1:1000, Abcam; Cambridge, MA, U.S.A., ab48187), IRE1-a (1:1000, Abcam; Cambridge, MA, U.S.A., ab37073), CIRP (1:5000, Proteintech Biotechnology, WH, China, 10209-2-AP), CSE (1:1000, Proteintech Biotechnology, WH, China, 60234-1-Ig), and GAPDH (1:5000, Proteintech Biotechnology, WH, China, 10494-1-AP).

Techniques: Western Blot, Control, Staining

Journal: Bioscience Reports

Article Title: Hydrogen sulfide attenuates sepsis-induced cardiac dysfunction in infant rats by inhibiting the expression of cold-inducible RNA-binding protein

doi: 10.1042/BSR20241398

Figure Lengend Snippet: ( A−F ) Representative Western blots and quantitative analysis for GRP78, ATF6, p-PERK, PERK, p-IRE1-a, IRE1-a. GAPDH was used as the internal control. ( G ) Representative H&E staining left ventricular sections (scale bar = 50μm μm). ( H ) Severity scores of heart tissue sections. Data are mean ± SD from four independent experiments ( n = 6). One-way analysis of variance followed by LSD test with equal variance, and the Dunnett’s T3 method was performed if there was missing variance, P ＜0.05 was considered significant vs. sham + V. H&E, hematoxylin and eosin.

Article Snippet: The membranes were blocked with 3% BSA for 1.5 h at room temperature, and antigens were detected using the following antibodies at 4°C overnight: GRP78 (1:10000, Santa Cruz Biotechnology Inc., TX, U.S.A., sc-166490), ATF6 (1:500, MCE, SH, China, HY- P80379 ), p-PERK (1:1000, GeneTex, SZ, China, GTX00673), PERK (1:1000, Santa Cruz Biotechnology, TX, U.S.A., sc-377400), p-IRE1-a (1:1000, Abcam; Cambridge, MA, U.S.A., ab48187), IRE1-a (1:1000, Abcam; Cambridge, MA, U.S.A., ab37073), CIRP (1:5000, Proteintech Biotechnology, WH, China, 10209-2-AP), CSE (1:1000, Proteintech Biotechnology, WH, China, 60234-1-Ig), and GAPDH (1:5000, Proteintech Biotechnology, WH, China, 10494-1-AP).

Techniques: Western Blot, Control, Staining

Journal: Bioscience Reports

Article Title: Hydrogen sulfide attenuates sepsis-induced cardiac dysfunction in infant rats by inhibiting the expression of cold-inducible RNA-binding protein

doi: 10.1042/BSR20241398

Figure Lengend Snippet: CIRP and marker proteins amount of endoplasmic reticulum stress was significantly decreased after NaHS treatment:( A-–G ) Representative Western blots and quantitative analysis for CIRP, GRP78, ATF6, p-PERK, PERK, p-IRE1-a, and IRE1-a. GAPDH was used as the internal control. ( H ) Representative H&E staining left ventricular sections (scale bar = 50 μm). ( I ) Severity scores of heart tissue sections. Data are mean ± SD from four independent experiments ( n = 6). One-way analysis of variance followed by LSD test with equal variance, and the Dunnett’s T3 method was performed if there was missing variance, P ＜0.05 was considered significant vs. Ssham + V. H&E, hematoxylin and eosin.

Article Snippet: The membranes were blocked with 3% BSA for 1.5 h at room temperature, and antigens were detected using the following antibodies at 4°C overnight: GRP78 (1:10000, Santa Cruz Biotechnology Inc., TX, U.S.A., sc-166490), ATF6 (1:500, MCE, SH, China, HY- P80379 ), p-PERK (1:1000, GeneTex, SZ, China, GTX00673), PERK (1:1000, Santa Cruz Biotechnology, TX, U.S.A., sc-377400), p-IRE1-a (1:1000, Abcam; Cambridge, MA, U.S.A., ab48187), IRE1-a (1:1000, Abcam; Cambridge, MA, U.S.A., ab37073), CIRP (1:5000, Proteintech Biotechnology, WH, China, 10209-2-AP), CSE (1:1000, Proteintech Biotechnology, WH, China, 60234-1-Ig), and GAPDH (1:5000, Proteintech Biotechnology, WH, China, 10494-1-AP).

Techniques: Marker, Western Blot, Control, Staining

Journal: Frontiers in Immunology

Article Title: ß1-adrenergic blockers preserve neuromuscular function by inhibiting the production of extracellular traps during systemic inflammation in mice

doi: 10.3389/fimmu.2023.1228374

Figure Lengend Snippet: Antibodies used for cell processing before flow cytometry analyses.

Article Snippet: TFAM Secondary antibody goat anti rabbit , Boster (PB9447) (Abcam Cat# ab130805, RRID : AB_11156672) , Unconjugated APC.

Techniques: Flow Cytometry