sh sy5y human neuroblastoma cell lines  (ATCC)

Journal: Alzheimer's & Dementia

Article Title: NRN1 as a therapeutic target for Alzheimer's disease

doi: 10.1002/alz.71149

Figure Lengend Snippet: Abcam ab64186 recognizes NRN1 protein in mouse neuroblastoma cells. (A) Representative Western blot of human embryonic kidney (HEK) 293T, Neuro‐2a (N2a) mouse neuroblastoma, and SH‐SY5Y human neuroblastoma cell lysates (50 µg protein), probed with NRN1 polyclonal antibody Abcam ab64186 and GAPDH monoclonal antibody. (B) N2a cells were transfected with NRN1 or Scramble (non‐targeting) siRNA smart pools and harvested after 96 h. Western blot analyses with NRN1 polyclonal antibody Abcam ab64186 revealed reduced intensity of the ∼34 kDa band in NRN1‐depleted cells, compared to Scramble control. 10 µg of protein was loaded per lane, and GAPDH was probed as a loading control. GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase; NRN1, Neuritin‐1; siRNA, small interfering RNA;

Article Snippet: SH‐SY5Y human neuroblastoma cell lines (Catalog No.: CRL‐2266, ATCC) were maintained in a 1:1 mixture of DMEM/F12 (Catalog No.: 11330057, Thermo Fisher Scientific) and Eagle's Minimum Essential Medium (Catalog No.: 50‐238‐2632, Fisher Scientific) with 10% FBS and 1% penicillin‐streptomycin.

Techniques: Western Blot, Transfection, Control, Small Interfering RNA

Journal: Journal of Neurochemistry

Article Title: Prosaposin Is Cleaved Into Saposins by Multiple Cathepsins in a Progranulin‐Regulated Fashion

doi: 10.1111/jnc.70357

Figure Lengend Snippet: Progranulin and multi‐granulin fragments influence the rate of PSAP cleavage and production of saposins (A–D). (A) Model of progranulin (PGRN) showing full length, and the three multi‐granulin fragments (MGFs) pG, BAC, and CDE. *Indicates the PSAP binding site on PGRN. (B–E) PSAP was incubated at 37°C for a 6 h time course with CTSD (400 nM) and full‐length PGRN, pG, BAC, or CDE (400 nM) at pH 4.5. (F–I) PSAP was incubated with CTSD and CDE as in (B–E) but a shorter time course of 30 min. Samples were run on western blot and probed with α‐SapA, α‐SapB, α‐SapC, α‐SapD antibodies. (J) Schematic of lysosome isolation from SH‐SY5Y neuronal culture by LysoIP (Biorender). (K) PSAP was incubated at 37°C for up to 6 h with SH‐SY5Y LysoIP lysosomes (12.5 ng/μL) with or without CDE (400 nM) at pH 4.5. Data represents three independent replicates.

Article Snippet: SH‐SY5Y human neuroblastoma cells were obtained from American Type Culture Collection (ATCC, CRL‐2266).

Techniques: Binding Assay, Incubation, Western Blot, Isolation