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. 2020 Jan 21;29(6):892–906. doi: 10.1093/hmg/ddaa010

Figure 4.

Figure 4

Calpain-1 knockout counteracts excessive cleavage of synaptic proteins in MJD animals. (A) Western blot of synapsin-1 and synapsin-2a in whole brain of 3-month-old mice. Actin served as a loading control. (B) Densitometric quantification of synapsin-1 and -2a full-length levels and fragmentation (WT/Capn1+/+: n = 3; WT/Capn1−/−: n = 3; 84Q/Capn1+/+: n = 2; 84Q/Capn1−/−: n = 3). *(black): WT/Capn1+/+ versus WT/Capn1−/−, *(blue): 84Q/Capn1+/+ versus 84Q/Capn1−/−, #:WT/Capn1+/+ versus 84Q/Capn1+/+; */#P ≤ 0.05, **/##P ≤ 0.01, ###P ≤ 0.001, ****P ≤ 0.0001; one-way ANOVA with Fisher’s LSD post-test. (C) Western blot of SNAP-25 and PSD-95 in whole brain of 3-month-old mice. Vinculin served as a loading control. (D) Densitometric quantification of SNAP-25 cleavage and PSD-95 levels (WT/Capn1+/+: n = 3; WT/Capn1−/−: n = 3; 84Q/Capn1+/+: n = 2; 84Q/Capn1−/−: n = 3). *(black): WT/Capn1+/+ versus WT/Capn1−/−, *(blue): 84Q/Capn1+/+ versus 84Q/Capn1−/−, #:WT/Capn1+/+ versus 84Q/Capn1+/+; *P ≤ 0.05, **P ≤ 0.01, ###P ≤ 0.001, ****P ≤ 0.0001; one-way ANOVA with Fisher’s LSD post-test. White arrowheads indicate full-length proteins, black arrowheads protein fragments, ‘a’ and ‘b’ mark respective synapsin isoforms. fl = full-length, LE = long exposure, SE = short exposure. Columns represent means.