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. 2022 Aug 22;23(16):9489. doi: 10.3390/ijms23169489

Figure 4.

Figure 4

Generation of doxycycline–inducible HEK293 and INS1 cell lines for AP–MS. (A) Schematic pipeline of the AP-MS. Signal peptide (PRE), StrepII, and HA tags are indicated. (B) CDNF expressed from pre-SH-CDNF construct is biologically active. Nerve growth factor (NGF)-maintained mouse embryonic SCG neurons were injected with the indicated expression plasmids and treated with 2 µM tunicamycin. The number of surviving injected neurons was expressed as % of the number of living injected neurons counted 3–4 h after injection. Shown are the means of five independent experiments ± S.E.M. Data of each experimental group were compared to respective empty vector control using ordinary ANOVA and Sidak’s multiple comparison post hoc test. **** denotes p < 0.0001. The null hypothesis was rejected at p < 0.05. (C) Expression of SH-tagged CDNF upon doxycycline induction in Hek293 and INS1 cell lines. Whole-cell lysates of HEK293 and INS1 parental and CDNF-expressing (SH-CDNF) cell lines were analyzed by immunoblotting using the indicated antibodies. Recombinant human CDNF was used as an antibody control.