Figure 4.

cGAS and STING pharmacological inhibitors ameliorates ALS neuropathology in the SOD1-G93A model

(A and B) SIM-A9 cells co-transfected with luciferase reporters and 0.01 μg of the indicated SOD1 mutant plasmids in the presence of RU.521, H-151, or C-176. Estimated EC₅₀ values from dose-response curves of the indicated inhibitors were measured.

(C–E) Kaplan-Meier plot of disease-onset days, survival rate, or motor performance of indicated time-point for G93A, Sting^−/−, or G93A/Sting^−/−-double-positive mice. Schematic diagram indicates how the gait performance (stride-length, blue line) of each mouse was measured.

(F) In vivo ChAT and LDH levels from brain and spinal cord homogenates isolated at the indicated time-points in G93A, Sting^−/−, or G93A/Sting^−/−-double-positive mice.

(G) Expression of the indicated mRNA in vivo in brain isolated from week-21 of G93A, Sting^−/−, or G93A/Sting^−/−-double-positive mice.

(H–J) Body weight of vehicle and inhibitor-treated groups were measured (H). Kaplan-Meier plot of disease-onset days, or survival time (I and J) of vehicle and inhibitor-treated groups. Number of mice used in each group as indicated.

(K) Grip strength, hindlimb grasping, or gait performance test of G93A mice after challenge with vehicle, RU.521 (500 μg), or C-176 (500 μg).

(L and M) Basal gene expression of indicated mRNA in vivo in brain and spinal cord isolates from G93A mice challenged with vehicle, RU.521 (500 μg, i.p.), or C-176 (500 μg, i.p.) at Week 21 analyzed by RT-qPCR. Number of dots = number of mice (biological replicates) used in each condition. All RT-qPCR of RNA samples was normalized to β-actin. Unless indicated, data represent the mean of at least three independent experiments (mean ± SEM). ∗p< 0.05, ∗∗p< 0.01, ∗∗∗p< 0.001, ∗∗∗∗p< 0.0001; (E–H and K–M: unpaired Student’s t test; C, D, I, and J: Log rank Mantel-Cox test).