Ab85670

Antibodies used in the study were as follows: anti-Flag (F3165/F7425, Sigma), anti-Myc (9B11/71D10, Cell Signaling Technology (CST)), anti-HA (16B12, Biolegend and H6908, Sigma), anti-TRAF2 (sc-136999, Santa Cruz), anti-Rab5 (sc-46692, Santa Cruz), anti-Rab7 (sc-376362, Santa Cruz), anti-Sprouty 2 (sc-100862, Santa Cruz/ab85670, Abcam), anti-EGFR (sc-373746, Santa Cruz), anti-p-Tyr (9411, CST), anti-p-Ser/Thr (ab9344, Abcam and 05-368, Sigma), anti-Ser (sc-81514, Santa Cruz), anti-Thr (sc-5267, Santa Cruz), anti-Ubiquitin (sc-8017, Santa Cruz), anti-β-Actin (AC026, Abclonal), anti-GFP (AE011, Abclonal), anti-glutathione S-transferase (GST) (2622 S, CST), anti-V5 (R960-25, Thermo Fisher and 30801ES10, Yeasen), anti-Mouse/Rabbit IgG-peroxidase secondary antibody (A0545/A9044, Sigma), anti-Mouse IgG (light chain specific)-peroxidase secondary antibody (115-005-174, Jackson), and anti-DYRK1A polyclonal antibody has been previously described [30 (link)].

The hippocampal tissues of the mice from different groups were treated with normal saline, fixed with 4% paraformaldehyde for 24 h, treated with 0.1 M PBS and subsequently dehydrated in graded ethanol after being embedded in paraffin. An immunofluorescence assay was then performed on 3.5 μm-thick coronal sections. Following retrieval, the sections were incubated in 0.1 M PBS containing 10% serum for 40 min and incubated overnight at 4 °C with primary antibodies: mouse monoclonal antibody [Mec-168] against MeCP2 (ab50005, 1:1000, Abcam, Inc., Cambridge, UK) and rabbit polyclonal antibodies against SPRY2 (ab85670, 1:1000, Abcam, Inc., Cambridge, UK) and NeuN (mouse 1:1,000; 2,742,283, Millipore, Billerica, MA, USA). After three 0.1 M PBS washes (5 min per wash), the sections were incubated with fluorescein isothiocyanate (FITC)-conjugated secondary goat anti-rabbit immunoglobulin G (IgG) (1:100; SA00003-2, Proteintech Group, Chicago, IL, USA) or tetramethylrhodamine isothiocyanate-conjugated anti-mouse IgG (1:100; SA00007-1, Proteintech Group, Chicago, IL, USA) at room temperature for 4 h. The sections were counterstained with 4’,6-diamidino-2-phenylindole for 5 min. Immunofluorescence images were obtained using a Nikon Eclipse NI microscope.