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. 2023 Sep 8;10(31):2301120. doi: 10.1002/advs.202301120

Figure 1.

Figure 1

Effects of different intrabody fragments on mHTT. a) Schematics of the intrabody fragments SM1, SM2, and SM3 (24, 29, and 23 amino acids), and N‐terminal mutant HTT fragment (HTT‐N171‐150Q). The intrabody peptides are overlapping fragments of variable region heavy (VH) chain of the scFv‐mEM48, which is tagged with the HA epitope and linked to a LAMP1 signal. N‐terminal mHTT fragment (HTT1–171aa) contains 150 glutamine repeats (150Q). b) Transfection of SM1, SM2, or SM3 with N171‐150Q in HEK 293T cells showed that SM3 can reduce aggregates more efficiently. Scale bar: 20 µm. c) Western blotting analysis of SM1 or SM3 and mutant HTT transfected HEK293T cells using mEM48 antibody. d) Quantification of the aggregates in co‐transfected HEK 293T cells (n = 8 images per group). Data were analyzed by one‐way ANOVA with Dunnett's multiple comparisons test and are presented as mean ± SEM. **P = 0.0085 (150Q+HA vs 150Q+SM1); **P = 0.0020(150Q+HA vs 150Q+SM2); ****P < 0.0001 (150Q+HA vs 150Q+SM3). e) Quantification of the ratios of mHTT to vinculin on the western blots. The data were obtained from four independent experiments (n = 4). Data were analyzed by one‐way ANOVA with Dunnett's multiple comparisons test and presented as mean ± SEM. **P = 0.0025; ***P = 0.0003; ****P < 0.0001. f) Quantification of HEK293 cell viability using cell counting kit‐8 (CCK8). The data were obtained from four independent experiments (n = 4). Data were analyzed by one‐way ANOVA with Dunnett's multiple comparisons test and presented as mean ± SEM. *P = 0.031; ***P = 0.0001.