Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: The high expression of circ-ACTR2 was detected in DN samples and HG-treated HRMCs. A , B The qRT-PCR was performed to detect the circ-ACTR2 level in normal and DN samples ( A ) or control and HG-treated HRMCs ( B ). C The determination of circ-ACTR2 and linear ACTR2 was conducted by qRT-PCR after treatment of RNase R in total RNA. D The circ-ACTR2, U6 and GAPDH levels were assayed by qRT-PCR in cytoplasm and nucleus. ** P < 0.01, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: HG-induced cell damages in HRMCs were partly reversed by the silence of circ-ACTR2. A The expression of circ-ACTR2 was quantified using qRT-PCR in four groups of control, HG, HG + si-NC, HG + si-ACTR2 in HRMCs. B Cell viability detection was performed by CCK-8 assay. C Inflammatory cytokines IL-6 and TNF-α were determined by ELISA. D Cell proliferation analysis was performed by EdU assay. E The protein examination of collagen I and collagen IV was conducted by western blot. F , G SOD activity ( F ) and MDA level ( G ) were respectively measured using the corresponding kits. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Quantitative RT-PCR, Control, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, EdU Assay, Western Blot, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2 exhibited the sponge function of miR-205-5p. A Starbase v2.0 predicted the binding sites between circ-ACTR2 and miR-205-5p. B The miR-205-5p level was examined by qRT-PCR after transfection of miR-NC or miR-205-5p. C , D Dual-luciferase reporter assay ( C ) and RIP assay ( D ) were used to confirm whether circ-ACTR2 combined with miR-205-5p. E The quantification of miR-205-5p was carried out using qRT-PCR in normal and DN tissues. F The linear analysis between circ-ACTR2 and miR-205-5p was performed using the Pearson’s correlation coefficient. G The effect of HG on the miR-205-5p expression was analyzed via qRT-PCR. H The level of circ-ACTR2 was detected by qRT-PCR in control, HG, HG + pcD5-ciR or HG + circ-ACTR2 group. I The regulation of circ-ACTR2 inhibition or overexpression on the miR-205-5p expression was performed by qRT-PCR in HG-treated HRMCs. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Binding Assay, Quantitative RT-PCR, Transfection, Luciferase, Reporter Assay, Expressing, Control, Inhibition, Over Expression

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2/miR-205-5p axis affected the regulation of HG in HRMCs. A The transfection efficiency of anti-miR-205-5p was assessed by qRT-PCR. B The qRT-PCR was applied for the expression detection of miR-205-5p after transfection of si-NC, si-ACTR2, si-ACTR2 + anti-miR-NC or si-ACTR2 + anti-miR-205-5p in HG-treated HRMCs. C Cell viability was examined via CCK-8 assay. D The concentrations of IL-6 and TNF-α were tested via ELSIA. E Cell proliferation was assessed via EdU assay. F , G The levels of collagen I and collagen IV were assayed via western blot. H , I Oxidative stress was evaluated by SOD activity ( H ) and MDA level ( I ) using the kits. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Transfection, Quantitative RT-PCR, Expressing, CCK-8 Assay, EdU Assay, Western Blot, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: HMGA2 downregulation protected against the HG-induced cell dysfunction in HRMCs. A The protein expression of HMGA2 in control, HG, HG + si-NC or HG + si-HMGA2 group was evaluated by western blot. B CCK-8 was performed to analyze cell viability. C ELISA was performed to measure the concentrations of IL-6 and TNF-α. D EdU assay was conducted to determine cell proliferation. E , F Western blot was conducted to detect the protein levels of collagen I and collagen IV. G , H SOD activity ( G ) and MDA level ( H ) by the detection kits were exploited to examine the oxidative stress. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Expressing, Control, Western Blot, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, EdU Assay, Activity Assay

Journal: Diabetology & Metabolic Syndrome

Article Title: Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

doi: 10.1186/s13098-021-00692-x

Figure Lengend Snippet: Circ-ACTR2 regulated the HMGA2 level by acting as a sponge of miR-205-5p. A , B The mRNA ( A ) and protein ( B ) levels of HMGA2 were respectively detected by qRT-PCR and western blot after transfection of si-NC, si-circ-ACTR2, si-circ-ACTR2 + anti-miR-NC or si-circ-ACTR2 + anti-miR-205-5p in HG-treated HRMCs. ** P < 0.01, **** P < 0.0001

Article Snippet: Human renal mesangial cell line HRMC (BioVector NTCC Inc., Beijing, China) was maintained in cell medium produced by Dulbecco’s modified eagle medium (DMEM; Hyclone, Logan, UT, USA), 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% antibiotics (Sigma-Aldrich).

Techniques: Quantitative RT-PCR, Western Blot, Transfection

Journal: Molecular Oncology

Article Title: Obesity alters the fitness of peritumoral adipose tissue, exacerbating tumor invasiveness in renal cancer through the induction of ADAM12 and CYP1B1

doi: 10.1002/1878-0261.13782

Figure Lengend Snippet: Adipose peritumoral tissue affects the expression of inflammatory genes by renal cell carcinoma (RCC) cells. (A) Relative mRNA expression of the different tumor suppressor genes in the healthy and tumor kidney tissues of RCC patients ( n = 10), including VHL , PBRM1 , SETD2 , and BAP1 , was measured by real‐time PCR. Cyclophilin‐A was used as an internal control. Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by Student's t ‐test. (B) Schematic figure illustrating the protocol for differentiation of adipocytes from peritumoral adipose tissue (AT) of lean, overweight, and obese RCC patients. (C) Relative mRNA expression of the leptin and adiponectin of the differentiated adipocytes among the lean, overweight, and obese groups ( n = 6). Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by one‐way ANOVA. (D) Schematic figure of the conditioned medium (CM) experiment, showing RCC cells treated with conditioned media (CMs) of lean, overweight, and obese differentiated adipocytes. (E) Relative mRNA expression of the IL6 , CXCR4 , SDF1 , and BAFFR of the A498 cells treated with CMs collected from differentiated adipocytes derived from the lean, overweight, and obese groups ( n = 6). Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by one‐way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The human renal cancer cell line (A498) was grown in DMEM (Microgem); supplemented with 10% FBS, 1% Pen/Strep, and 1% L‐Glutamine, at 37 °C in 5% CO 2 .

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Derivative Assay