Figures and data
Normalized downstroke time
(A), decay time (B), and beating rate as assessed by beats per minute (C) in hiPSC-CM treated with bovine serum albumin (BSA), (control), TNF-α, IFN-γ, TNF-α plus mitoTempo, and IFN-γ plus mitoTempo. N=5-14. Data were analyzed by ordinary one-way ANOVA and post-hoc Tukey’s multiple comparison test. Bars represent group mean.
(A) Representative image of hiPSC-CM. Scale bar represents 100 μm. (B) Representative curves of calcium transient and its derivative in hiPSC-CM. (C) MMP as assessed by tetramethylrhodamine ethyl ester (TMRE) in hiPSC-CM treated with various cytokines. Concentration of cytokines used is as follows: CCL3 1nM, TNF-α 300 pg/ml, IL-6 1 ng/ml, IL-1β 200 pg/ml, IP-10 (or CXCL10) 500 pg/ml, IFN-γ 1ng/ml. N=4, Data were analyzed by ordinary one-way ANOVA and post-hoc Tukey’s multiple comparison test. Bars represent group mean.
OCR in hiPSC-CM treated with cytokines and antioxidant agent NAC.
(A) OCR trace of hiPSC-CM treated with BSA, TNF-α, IFN-γ, TNF-α+IFN-γ, TNF-α+NAC, IFN-γ+NAC, and TNF-α+IFN-γ+NAC. (B-D) Bar graph summary of data in Panel A with OCR at baseline (B), after adding carbonyl cyanide m-chlorophenyl hydrazone (CCCP) (C), and after adding antimycin A and rotenone (D). N=5-6. Data were analyzed by ordinary one-way ANOVA and post-hoc Tukey’s multiple comparison test. Bars represent group mean.
Decay time in hiPSC-CM after treatment with TNF-α and various ART drugs.
(A) Decay time after treatment with TNF-α, tenofovir, and TNF-α+tenofovir. (B) Decay time after treatment with TNF-α, darunavir, and TNF-α+darunavir. (C) Decay time after treatment with TNF-α, raltegravier, and TNF-α+ raltegravier. (D) Decay time after treatment with TNF-α, emtricitabine, and TNF-α+ emtricitabine. N=5-21. Data were analyzed by ordinary one-way ANOVA and post-hoc Tukey’s multiple comparison test. Bars represent group mean.
Cell viability (A) and cellular ROS levels (B) in hiPSC-CM exposed to concentrations ranging from between 3nM and 10μM of tenofovir (a nucleotide-analog reverse transcriptase inhibitor), darunavir (a protease inhibitor), raltegravir and elvitegravir (integrase inhibitors).
Decay time in hiPSC-CM after treatment with TNF-α and riociguat, sildenafil citrate, PF-04447943, and dapagliflozin.
(A) Decay time after treatment with TNF-α, sGS agonist riociguat, and TNF-α+riociguat. (B) Decay time after treatment with TNF-α, sildenafil citrate, and TNF-α+sildenafil citrate. (C) Decay time after treatment with TNF-α, PF-04447943, and TNF-α+PF-04447943. (D) Representative curve of calcium transient in hiPSC-CM treated with TNF-α, dapagliflozin, and TNF-α+dapagliflozin. (E) Curve of derivative calculated with data in panel (D). (F) Decay time after treatment with TNF-α, dapagliflozin, and TNF-α+dapagliflozin. (G) Downstroke time after treatment with TNF-α, dapagliflozin, and TNF-α+dapagliflozin. (H) Representative curve of calcium transient in hiPSC-CM treated with IFN-γ, dapagliflozin, and IFN-γ+dapagliflozin. (I) Curve of derivative calculated with data in panel (H). (J) Decay time after treatment with INF-γ, dapagliflozin, and IFN-γ+dapagliflozin. (K) Downstroke time after treatment with INF-γ, dapagliflozin, and IFN-γ+dapagliflozin. (N=5-21. Data were analyzed by ordinary one-way ANOVA and post-hoc Tukey’s multiple comparison test. Bars represent group mean.
Decay time (A) and downstroke time (B) in hiPSC-CM after treatment with TNF-α and 1 μM of dapagliflozin. N = 5-21, Data were analyzed by ordinary one-way ANOVA and post-hoc Tukey’s multiple comparison test. Bars represent group mean.
Ca2+-deay time of hiPSC-CM and angiogenic function of ECs after treatment with serum from Group 1 HIV+ patients with DD.
(A-C) Pooled normalized decay time (A), downstroke time (B) and beating rate (C) in hiPSC treated with 10% serum for 48 hours from group 1 HIV+ patients with DD. (D-G) Angiogenic parameters, including total length of branching (D), (N = 3), number of junctions (E) (N = 3), total length (F) (N =3) and number of meshes (G) (N = 3) in HUVECs after treatment with 2% serum from Group 1 patients for 20 hours. Data were analyzed by unpaired Student’s t test. Bars represent group mean.
Representative images of formed tube structure with HUVECs treated with 2% serum from HIV+ patients with high or low MPR for 20 hours.
Representative magnified images of formed tube structure with HUVECs treated with 2% serum from HIV+ patients with high or low MPR for 20 hours.
Diastolic function of hiPSC-CMs and angiogenic function of ECs after treatment with serum from HIV+ patients with DD (Group 2).
(A-B) Individual normalized decay time 3 hours (N=3-9) (A) or 24 hours (N=3-13) (B) after treatment of hiPSC-CM with 2% serum from patients. (C-D) Pooled decay time data from patients in panels A and B with 3 hour (C) and 24 hours (D) after treatment with serum. (E) Representative image of formed tube structure with cultured HUVECs treated with serum from HIV+ patients with DD. (F-I) Assessment of tube formation of HUVECs 20 hours after treatment with serum of patients, including number of meshes (F) (N = 8), number of junctions (G) (N = 8), number of segments (H) (N = 8) and total length of branching (I) (N = 8). Data were analyzed by unpaired Student’s t test for (C, D, F, G, H and I). Bars represent group mean.
