Journal: Stem Cell Research & Therapy

Article Title: Evaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drug

doi: 10.1186/s13287-022-02827-x

Figure Lengend Snippet: Effect of drugs on gene expression in photoreceptor-directed fibroblasts derived from an EYS-RP patient, Pt#1. The fibroblasts of a healthy individual (N#3) and fibroblasts of a patient suffering from retinitis pigmentosa due to a homozygous mutation in the EYS gene (Pt#1) were transdifferentiated to photoreceptor-like cells by retroviral transduction of four transcription factors. Gene expression was compared 2 weeks post-transduction. Differentiation medium was supplemented with four drugs, 4-phenylbutyric acid (4-PBA), metformin (Metf), rapamycin (Rapa), N-acetyl-L-cysteine (NAC) or the vehicle (water and ethanol (EtOH)) or none (-). The gene expression with each pharmacological treatment was compared to the gene expression in photoreceptor-like cells derived from fibroblasts of healthy individual without any supplement (-). Columns represent mean ± SEM. All data points are overlaid ( n = 6 (N#3 without any supplement (-)), n = 3 (N#3 with supplement and Pt#1). a = p > 0.05, b = p > 0.01, c = p > 0.001; One-way ANOVA followed by Tukey’s honest test. For the sake of simplicity, comparison with the gene expression level in N#3 without any supplement (-) is shown here. Comparison among other groups is shown in Additional file : Fig. S2

Article Snippet: After 5–6 h of retroviral transduction, the media in transduced fibroblast cells were replaced with differentiation media containing either one of the four drugs (rapamycin (Selleck chemicals, Cat#S1039), 4-PBA (WAKO, Cat#168-06471), NAC (WAKO, Cat#015-05132), metformin (Abcam, Cat#ab120847)), vehicle or no addition for Pt#1 and containing vehicle or no addition for N#3.

Techniques: Expressing, Derivative Assay, Mutagenesis, Transduction

Journal: PLoS ONE

Article Title: Chemical proteasome inhibition as a novel animal model of inner retinal degeneration in rats

doi: 10.1371/journal.pone.0217945

Figure Lengend Snippet: (A) Vehicle (open column, 10–100% DMSO in distilled water), MG-262 alone (closed column, 0.1 nmol/eye) and combined with the NMDA receptor antagonist memantine (dark grey, 100 nmol/eye), the calcium channel blocker lomerizine (light grey, 50 nmol/eye), the serine protease inhibitor nafamostat (dark grey, 10 nmol/eye), the JNK inhibitor AS601245 (light grey, 10.5 nmol/eye), the p38 inhibitor SB239063 (dark grey, 20 nmol/eye), the ERK inhibitor U0126 (dark grey, 50 nmol/eye), the ER stress modulator salubrinal (light grey, 50 nmol/eye) or sodium 4-phenyl butyrate (PBA, dark grey, 100 nmol/eye). (B) Vehicle (open column, 30–33% DMSO in distilled water), 17-DMAG alone (closed column, DMAG, 25 nmol/eye) and combined with memantine (dark grey, 100 nmol/eye), lomerizine (light grey, 50 nmol/eye), nafamostat (hatched, 10 nmol/eye), AS601245 (dark grey, 10.5 nmol/eye), salubrinal (light grey, 12.5 nmol/eye) or PBA (hatched, 100 nmol/eye). Each pharmacological agent was premixed with a proteasome inhibitor and concurrently administered to animals. Following intravitreal injection, the retina was isolated and NFL gene expression was determined by real time PCR. The NFL gene expression level was normalized to that of GAPDH in each retinal sample and shown as the value relative to the respective control. Each value represents the mean ± S.E.M. of 5 to 10 eyes from 3 to 5 animals. Note that NFL downregulation by MG-262 and DMAG alone was statistically significant compared with the respective control group by Tukey’s multiple comparison test.

Article Snippet: N-acetyl cysteine (NAC), U0126 and sodium 4-phenyl butyrate (PBA, Wako Pure Chemical Industries, Ltd., Osaka, Japan), memantine (Sequoia, Pangbourne, UK), lomerizine (Organon, Roseland, NJ, USA), nafamostat (Yick-Vic Chemicals & Pharmaceuticals, Hong Kong), salubrinal and KB-R7943 (Tocris, Avenmouth, Bristol, UK), Z-VAD-FMK (ZVAD), SB-216763 and ansatrienin A (Enzo Life Sciences, Farmingdale, NY), UBEI-41 (4 [4-(5-Nitro-furan-2-ylmethylene)-3,5-dioxo-pyrazolidin-1-yl]-benzoic acid ethyl ester, Biogenova, Potomac, MD), aldehyde dehydrogenase (ALDH, Fluka, Mexico city, Mexico), and brain-derived neurotrophic factor (BDNF, Millipore, Darmstadt, Germany) were provided by the respective chemical vendors.

Techniques: Protease Inhibitor, Injection, Isolation, Expressing, Real-time Polymerase Chain Reaction