Fig 1: Acetylated α-tubulin levels in SPAST HSP patient and control PBMCs. Flow cytometry was used to measure acetylated α-tubulin in overall PBMCs and PBMC cell sub-types. (A) The forward-scatter and side-scatter (FSC/SSC) gated plots represent the total mononuclear cells. (B) Live PBMCs were identified using a cell viability dye. Viable cells have a relatively lower fluorescence level of the viability dye and vice versa for non-viable cells. Gating was applied to identify the proportion of viable cells. (C) CD2, CD19, and CD14 cell surface antibodies were used to identify PBMC cell sub-types T cells, B cells and monocytes. The scatter plot shows CD14 monocytes and CD19 B cells separated using the surface antibodies. The proportion of live PBMCs (D) and the proportion of T cells (E), B cells (F) and monocytes (G) is shown for all HSP-SPAST patients and healthy controls. (H–J) Levels of acetylated α-tubulin in PBMCs were measured in patient and control PBMC samples. Scatter plot of an acetylated α-tubulin negative control sample (H) and representative control (I) and patient (J) samples are shown. (K) Histogram plot showing acetylated α-tubulin levels in representative patient and control PBMC samples. (L) Patient and control group level comparison based on acetylated α-tubulin geometric mean fluorescence intensity in all PBMCs. (M) Histogram plot showing acetylated α-tubulin levels in representative patient and control T cells. Patient and control group level comparison of T cells (N), B cells (O) and monocytes (P) based acetylated α-tubulin geometric mean fluorescence intensity. Mean ± SEM.