Journal: Heliyon

Article Title: Role of NLRP3 inflammasome activation in HCC cell progression.

doi: 10.1016/j.heliyon.2023.e19542

Figure Lengend Snippet: Fig. 1. NLRP3 was expressed in low levels in hepatocellular carcinoma (HCC). (A, B and C) Protein expression in nine HCC cell lines, including SMMC-7721, Bel-7402, HepG2, Sk-Hep-1, MHCC-97L, Hep3B, MHCC-97H, Bel-7404, and Huh7 cells detected using western blotting. (A) NLRP3 protein. (B) ASC and Pro-caspase-1 protein. (C) Pro-IL-1β and Cleaved-IL-18 protein. (D) NLRP3 expression in Bel-7402 and SMMC-7721 cells was detected using immunofluorescence staining. FITC-conjugated secondary antibody staining (green) and DAPI (blue) staining indicated the location of NLRP3 and DAPI, respectively. (E) Quantitation of A, B and C. Data are expressed as mean ± SEM (n = 3).

Article Snippet: Antibodies against NLRP3 (rabbit polyAb, ab214185; goat polyAb, ab4207) were obtained from Abcam (Cambridge, UK), while antibodies against ASC (10500-1-AP), Caspase-1 (22915-1-AP), Pro-IL-1β (16806-1-AP), IKKβ (15649-1-AP), NF-κB (14220-1-AP), β-actin (6008-1-Ig), Fluorescein (FITC)-conjugated Affinipure Donkey AntiGoat IgG (SA00003-3), and Coralite594-conjugated Donkey Anti-Rabbit IgG (SA00013-8) were obtained from Proteintech Group, Inc (Wuhan, China).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Quantitation Assay

Journal: bioRxiv

Article Title: Effects of Hydroxychloroquine and Azithromycin on iPSC-derived Cardiomyocytes: Considerations for the Treatment of COVID-19 Patients

doi: 10.1101/2021.08.19.456950

Figure Lengend Snippet: A, Two representative western blots showing the expression of Nav1.5 and Cx43 in iPSC-CMs under different drug treatment conditions. B, Quantitation of protein expression levels of Nav1.5 in iPSC-CMs under different conditions; N = 6 independent differentiations. C, Quantitation of protein expression of Cx43 in iPSC-CMs under different drug treatment conditions; N = 6 independent differentiations. D, Representative images showing immunostaining for Nav1.5 (green) and Cx43 (red) in iPSC-CMs under different drug treatment conditions. Cell nuclei are shown in blue (Hoechst). Statistical evaluation was performed using one-way ANOVA with Tukey’s multiple comparison test (** p < 0.01, and *** p < 0.001).

Article Snippet: For staining, cells were incubated with the following primary antibodies: anti-α-actinin, clone EA-53 (1:500; mouse monoclonal, IgG1, Sigma-Aldrich, 7811), anti-Nav1.5 (1:200; rabbit polyclonal, Alomone Labs, ASC-005), and anti-Cx43, clone 2 (1:1000; mouse monoclonal, IgG1, BD Biosciences, 610061) at 4°C overnight.

Techniques: Western Blot, Expressing, Quantitation Assay, Immunostaining

Journal: bioRxiv

Article Title: Effects of Hydroxychloroquine and Azithromycin on iPSC-derived Cardiomyocytes: Considerations for the Treatment of COVID-19 Patients

doi: 10.1101/2021.08.19.456950

Figure Lengend Snippet: A, Representative I Na traces of iPSC-CMs in the control and drug-treated groups (10 µM AZM, 10 µM HCQ, and their combination). B, Statistical analysis of membrane capacitance of iPSC-CMs in the control and drug-treated groups. Different shapes of symbols indicate different differentiations. C, Statistical analysis of I Na in control and 7-day drug-treated groups. D, E Steady-state activation (D) and inactivation (E) in control and 7-day drug treated groups. I Na steady-state kinetics are altered by AZM and HCQ. n=41, 40, 30, and 41 cells for the control, 10 µM AZM-treated, 10 µM HCQ-treated, and AZM and HCQ combination groups, respectively, were analyzed; shown are mean and SEM from 6 independent differentiations (B-E). One-way ANOVA with Tukey’s multiple comparison test (B) and two-way ANOVA with Bonferroni post-hoc test (C-E) were used for statistical analysis (* p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.001).

Article Snippet: For staining, cells were incubated with the following primary antibodies: anti-α-actinin, clone EA-53 (1:500; mouse monoclonal, IgG1, Sigma-Aldrich, 7811), anti-Nav1.5 (1:200; rabbit polyclonal, Alomone Labs, ASC-005), and anti-Cx43, clone 2 (1:1000; mouse monoclonal, IgG1, BD Biosciences, 610061) at 4°C overnight.

Techniques: Activation Assay