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Fig. 5 | Cell Communication and Signaling

Fig. 5

From: The extracellular HDAC6 ZnF UBP domain modulates the actin network and post-translational modifications of Tau

Fig. 5

Podosome, lamellipodia and podonut-like structure induced by HDAC6 ZnF UBP. Neuro2a cells exposed to HDAC6 ZnF UBP exhibit a variety of actin rich structures characteristics of migratory or invading cells. a–c) Untreated neuro2a cells showed actin uniformly distributed along the periphery with small neuritic extensions while d, e HDAC6 ZnF UBP treated cells showed longer extensions and membranes ruffles resembling podosomes involved in cell migration. f, g Invadopodia and lamellipodia-like structures (encircled) were also observed, which are found in phagocytic cells. h, i Most of the treated cells were observed to consist of assemblance of podosomes (encircled) and podosome clusters called podonuts (white arrow) rich in actin. j The percentage of cells with or without podosomes/neurite extensions was calculated for control and HDAC6 ZnF UBP treatments groups. There were < 20% cells in control with podosome-like structures while > 80% cells in exhibited podosome-like structures upon HDAC6 ZnF UBP treatments. In cell control, 65–70% cells had neurite extensions, which were calculated to be 80–85% in HDAC6 ZnF UBP treated cells. (P + and P- represent cells with and without podosomes respectively. N + and N- represent cells with or without neuritic extensions respectively.) k The overall neurite extensions in neuro2a cells were counted in untreated and HDAC6 treated group in β-actin immunostained cells. The number of extensions in HDAC6 treated cells were found to be greatly enhanced compared to untreated group. l The podosomes formed after HDAC6 ZnF UBP treatment were quantified by counting the podosome crown structures in both untreated and treated groups. HDAC6 ZnF UBP treated cells showed more podosome clusters signifying its actin modulating effect. Statistical significance determined by two-tailed unpaired t-test. (n.s.—non-significant, * indicates P ≤ 0.05, ** indicates P ≤ 0.01, *** indicates P ≤ 0.001)

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