Figure 1.
Loss of α4A- and β1-tubulins leads to severe spherocytosis and prevents marginal band formation. (A) Blood platelet counts of WT, KOA4A (A4A), KOB1 (B1), and DKO mice. Results are expressed in 109/L. Bar graph represents the mean ±SEM. Each data point corresponds to an individual mouse. N > 20 per strain. Statistical analysis was conducted by 1-way analysis of variance (ANOVA) followed by Tukey honest significant difference (HSD) post-hoc test. (B) Mean platelet volumes of WT, KOA4A (A4A), KOB1 (B1), and DKO mice. Results are expressed in fL. Bar graph represents the mean ±SEM. Each data point corresponds to an individual mouse. Statistical analysis was conducted by Welch ANOVA followed by Dunnett T3 post-hoc test. (C) Representative transmission electron micrographs of WT, KOA4A (A4A), KOB1 (B1), and DKO platelets. The top panels correspond to a large field of view. The lower panels correspond to a close-up view, and higher magnifications in the lower right corners show a cross-sectioned microtubule marginal band for WT, KOA4A, and KOB1 platelets that is absent in DKO platelets. Scale bar: 5 μm (top panel), 1 μm (bottom panel), and 250 nm (lower right corner). (D) Distribution of WT, KOA4A (A4A), KOB1 (B1), and DKO platelets according to their morphology. Platelets were manually ranked based on their morphology (disc, oval, or round). Results are expressed in % of each class. Bar graph (stacked) represents the mean ±SEM, with at least >100 platelets per genotype counted. Statistical analysis of the round morphology was conducted by 1-way ANOVA followed by Tukey HSD post-hoc test, with DKO as a reference. ∗∗∗P < .001. (E) Enzyme-linked immunosorbent assay quantification of total tubulin of platelet lysates of WT, KOA4A (A4A), KOB1 (B1), and DKO mice. Results are expressed in fg per platelet. Bar graph represents the mean ±SEM. Each data point corresponds to an individual platelet lysate, obtained from a single mouse. N > 3 per group. Statistical analysis was conducted by 1-way ANOVA followed by Tukey HSD post-hoc test.

Loss of α4A- and β1-tubulins leads to severe spherocytosis and prevents marginal band formation. (A) Blood platelet counts of WT, KOA4A (A4A), KOB1 (B1), and DKO mice. Results are expressed in 109/L. Bar graph represents the mean ±SEM. Each data point corresponds to an individual mouse. N > 20 per strain. Statistical analysis was conducted by 1-way analysis of variance (ANOVA) followed by Tukey honest significant difference (HSD) post-hoc test. (B) Mean platelet volumes of WT, KOA4A (A4A), KOB1 (B1), and DKO mice. Results are expressed in fL. Bar graph represents the mean ±SEM. Each data point corresponds to an individual mouse. Statistical analysis was conducted by Welch ANOVA followed by Dunnett T3 post-hoc test. (C) Representative transmission electron micrographs of WT, KOA4A (A4A), KOB1 (B1), and DKO platelets. The top panels correspond to a large field of view. The lower panels correspond to a close-up view, and higher magnifications in the lower right corners show a cross-sectioned microtubule marginal band for WT, KOA4A, and KOB1 platelets that is absent in DKO platelets. Scale bar: 5 μm (top panel), 1 μm (bottom panel), and 250 nm (lower right corner). (D) Distribution of WT, KOA4A (A4A), KOB1 (B1), and DKO platelets according to their morphology. Platelets were manually ranked based on their morphology (disc, oval, or round). Results are expressed in % of each class. Bar graph (stacked) represents the mean ±SEM, with at least >100 platelets per genotype counted. Statistical analysis of the round morphology was conducted by 1-way ANOVA followed by Tukey HSD post-hoc test, with DKO as a reference. ∗∗∗P < .001. (E) Enzyme-linked immunosorbent assay quantification of total tubulin of platelet lysates of WT, KOA4A (A4A), KOB1 (B1), and DKO mice. Results are expressed in fg per platelet. Bar graph represents the mean ±SEM. Each data point corresponds to an individual platelet lysate, obtained from a single mouse. N > 3 per group. Statistical analysis was conducted by 1-way ANOVA followed by Tukey HSD post-hoc test.

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