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Electrospinning provides a facile and versatile method for generating nanofibers from a large variety of starting materials, including polymers, ceramic, composites, and micro-/nanocolloids. In particular, incorporating functional nanoparticles (NPs) with polymeric materials endows the electrospun fibers/sheets with novel or better performance. This work evaluates the spinnability of polyacrylamide (PAAm) solution containing thermoresponsive poly(N-isopropylacrylamide-co-tert-butyl acrylate) microgel nanospheres (PNTs) prepared by colloid electrospinning. In the presence of a suitable weight ratio (1:4) of PAAm and PNTs, the infiber arrangements of PNTs-electrospun fibers will evolve into chain-like arrays and beads-on-string structures by confining of PAAm nanofibers, and then the free amide groups of PAAm can bind amide moieties on the surfaces of PNTs, resulting in the assembling of PNTs in the cores of PAAm fibers. The present work serves as a reference in the fabrication of novel thermoresponsive hybrid fibers involving functional nanospheres via electrospun packing. The prepared nanofibers with chainlike and thermoresponsive colloid arrays in the cores are expected to have potential application in various fields.