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. 2019 Feb 4;129(3):1047–1060. doi: 10.1172/JCI120572

Figure 5. Rescue of the actin polymerization defect in patients’ lymphocytes by drug-induced activation of the RhoA/ROCK pathway and by retroviral correction of ARHGEF1 expression.

Figure 5

(A) Representative FACS plots showing the effect of the RhoA activator II (RhoA act., 32 μg/ml for 1 hour) on actin polymerization in lymphocytes from PBMCs from a healthy donor and the patients (P1, P2). The experiment was performed 3 times. (B) Representative FACS analyses highlighting the effect of the ROCK inhibitor Y27632 (0.6 mg/ml for 1 hour) on the level of polymerized actin (F-actin) in T cell blasts from the patients and a healthy donor. The experiment was performed 3 times for P1 and twice for P2. (C) Representative confocal microscopy images showing the effect of the retroviral transduction of an ARHGEF1 construction (ARHGEF1-IRES-Thy1.1) or an empty vector (IRES-Thy1.1) on the level of F-actin in T cell blasts from P1 and a healthy donor. Original magnification, ×40 (48 hours after transduction). The experiment was performed twice. (D) Single-cell quantification of the fluorescence intensity of F-actin after ARHGEF1 expression in the confocal photographs shown in C. Between 80 and 130 cells were evaluated for each condition. ***P < 0.001, 2- tailed unpaired Student’s t test.