Figure 5
From: Systemic gene therapy with thymosin β4 alleviates glomerular injury in mice
Effect of exogenous TB4 on ADR-injured podocytes in vitro. (a) Conditionally immortalised mouse podocytes were treated with RPMI-1640/ADR/TB4 and analysed after 24 h. Expression of podocyte Tmsb4x (b) and Tmsb10 (c) mRNA after ADR/TB4 treatment (n = 5 independent experiments; one-way ANOVA with Tukey post hoc test). Effect of ADR/TB4 on (d) cell viability (n = 4 independent experiments; two-way ANOVA with Tukey post hoc test). Effect of ADR/TB4 on (e) podocyte cell area and (f) podocyte mean F-actin fluorescence (n = 5 independent experiments with 50 cells analysed per condition, per experiment; two-way ANOVA with Tukey post hoc test). (g) Representative images of podocytes stained with Acti-Stain™ 488 Phalloidin displaying cortical actin stress fibres, cytoplasmic stress fibres and unorganised actin fibres. Scale bar = 10 μm. Percentage of podocytes displaying (h) cortical actin stress fibres, (i) cytoplasmic stress fibres and (j) unorganised stress fibres (n = 5 independent experiments with 50 cells analysed per condition, per experiment; two-way ANOVA with Tukey post hoc test). Data are presented as mean ± SEM; *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001. TB4, Tmsb4x, thymosin β4; Tmsb10, thymosin β10; ADR, Adriamycin; IFN-ɣ, interferon-gamma; OD, optical density; GapDH, Glyceraldehyde 3-phosphate dehydrogenase; MTT, methyltetrazolium.
