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. Author manuscript; available in PMC: 2020 Dec 1.
Published in final edited form as: Neurobiol Dis. 2020 Oct 14;146:105130. doi: 10.1016/j.nbd.2020.105130

Figure 3. High Tau levels do not modulate TARDBP gene expression but regulate TDP-43 oligomerization in iHEK.

Figure 3.

(A) IB of TDP-43 Oligo01 in cytoplasmic and nuclear fractions of WT tau iHEK (−Tet and +Tet). (B) Relative density of HMW TDP-43 in the cytoplasmic fraction (−Tet vs. +Tet, *p=0.0253). (C) Relative density of HMW of TDP-43 in the nuclear fraction (−Tet vs. +Tet, ns, p=0.0756). (D) IB of TDP-43 Oigo01 in the cytoplasmic and nuclear fractions of P301L tau iHEK. (E) Relative density of HMW TDP-43 in the cytoplasmic fraction (−Tet vs. +Tet, ns, p=0.7002) of P301L tau iHEK. (F) Relative density of HMW TDP-43 in the nuclear fraction (ns, p=0.1863) of P301L tau iHEK. TDP-43-Oligo01 Ab was used to detect patterns in both compartments before and after Tet incubation. (G) IB of TDP-43 Oligo02 in the cytoplasmic and nuclear fractions of WT tau iHEK (−Tet and +Tet). (H) Relative density of HMW TDP-43 in the cytoplasmic fraction (−Tet vs. +Tet, ****p<0.0001) of WT tau iHEK. (I) Relative density of HMW TDP-43 in the nuclear fraction of WT tau iHEK (−Tet vs. +Tet, *p=0.0375). (J) IB of TDP-43 Oligo02 in the cytoplasmic and nuclear fractions of P301L tau iHEK (−Tet and +Tet). (K) Relative density of HMW TDP-43 in the cytoplasmic fraction (−Tet vs. +Tet; ns, p=0.0542) of P301L tau iHEK. (L) Relative density of HMW TDP-43 in the nuclear fraction of P301L tau iHEK (−Tet vs. +Tet, *p=0.0018). (M) IB of commercial TDP-43 Ab in the cytoplasmic and nuclear fractions of WT tau iHEK (−Tet and +Tet). (N) Relative density of HMW TDP-43 in the cytoplasmic fraction (−Tet vs. +Tet; ns, p=0.0793) of WT tau iHEK. (O) Relative density of HMW TDP-43 in the nuclear fraction of WT tau iHEK (−Tet vs. +Tet, **p=0.0069). (P) IB of commercial TDP-43 Ab in the cytoplasmic and nuclear fractions of P301L tau iHEK (−Tet and +Tet). (Q) Relative density of HMW TDP-43 in the nuclear fraction (−Tet vs. +Tet, ** p=0.0093) of P301L tau iHEK. (R-T) RT-qPCR of TARDBP expression in HEK-293, WT, and P301L-expressing tau iHEK cells, respectively. Unpaired t-tests were used to compared untreated (−Tet) and treated (+Tet) groups. HEK-293 −Tet vs. +Tet ns, p=0.4520; P301L tau iHEK −Tet vs. +Tet ns, p=0.3377 and WT tau iHEK ns, p=0.0588. The same membrane (for WT and P301L tau iHEK) was immunoblotted for three different Abs (TDP-43 Oligo01, TDP-43 Oligo02, and commercial TDP-43). Each membrane includes three samples per condition. Unpaired t-tests were performed to compare HMW TDP-43 in the cytoplasm and nucleus of −Tet and +Tet iHEK. Values are represented as box-and-whisker plots with bars (Mean±SD).