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Fig. 4 | Acta Neuropathologica Communications

Fig. 4

From: LMTK2 binds to kinesin light chains to mediate anterograde axonal transport of cdk5/p35 and LMTK2 levels are reduced in Alzheimer’s disease brains

Fig. 4

LMTK2, p35 and KLC1 form a complex. a Mutation of LMTK2(WD1388/1389AA) does not affect LMTK2 binding to p35. HEK293 cells were transfected with either control empty vector (Ctrl), myc-LMTK2, myc-LMTK2(WD/1388/1389AA), myc-LMTK2 + p35 or myc-LMTK2(WD1388/1389AA) + p35. LMTK2 was immunoprecipitated via the myc-tag and bound p35 detected by immunoblotting. Both inputs and immunoprecipitations (IP) are shown. b p35 forms a complex with cdk5, LMTK2 and KLC1, and mutation of LMTK2(WD1388/1389AA) specifically disrupts KLC1 binding to the complex. HEK293 cells were transfected with either control empty vector (Ctrl), HA-p35 + myc-LMTK2 + FLAG-KLC1 or HA-p35 + myc-LMTK2(WD1388/1389AA) + FLAG-KLC1. p35 was immunoprecipitated via the HA-tag and bound LMTK2, KLC1 and cdk5 detected on immunoblots. Both inputs and immunoprecipitations (IP) are shown. c LMTK2 mediates formation of a complex between KLC1 and p35 in cortical neurons. Neurons were either untreated or treated with control or LMTK2 siRNAs and PLAs for KLC1 and p35 performed using goat anti-KLC1 and rabbit anti-p35 antibodies. Neurons were also immunostained for tubulin to reveal architecture. KLC1-p35 signals are markedly reduced in both cell bodies and axons in LMTK2 siRNA treated neurons; arrows show PLA signals in axons. Graph shows quantification of PLA dots in axons in the different treated cells. Data were analysed by Welch’s ANOVA and Games-Howell post hoc test; N = 15–24, error bars are s.e.m., ***p < 0.001

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