Fig 1: Phase contrast and confocal imaging analysis of Hsd17b7 localization in differentiating Neuro2a cells. (A) Phase contrast images of cells grown in media supplemented with 10%FBS or 0.1%BSA for 24-h. Scale bar: 50 μM (B) Quantification of neurite structures (n > 300 cells) in 0.1% BSA (85 ± 5%) compared to the 10% FBS control group (11 ± 2%). Values in the graph are given as average ± standard error. *denotes p < 0.001. (C) 24-h post-transfection with Hsd17b7-GFP Neuro2a cells were serum deprived for an additional 24 h and co-stained with MAP2 antibody. Arrows denote GFP signal in MAP2-positive structures. Scale bar: 5 μM. (D) After 24-h serum deprivation, cells were stained for anti-Hsd17b7 antibody and co-stained with conjugated Phalloidin-488. Arrows indicate Hsd17b7 signal in Phalloidin positive neurite structures. Scale bar: 5 μM. (E) 24-h post-transfection with pCAG-GFP Neuro2a cells were serum deprived for an additional 24 h and co-stained with Hsd17b7 antibody. Arrows denote antibody signal in pCAG-GFP positive neurite structures. Scale bar: 5 μM.
Fig 2: Confocal images comparing endogenous Hsd17b7 vs. GFP-tagged Hsd17b7 in undifferentiated Neuro2a cells. (A,B) Neuro2a cells transfected with either an N- or C- terminally tagged Hsd17b7 GFP construct and (C) Undifferentiated Neuro2a cells stained for Hsd17b7 antibody. (D) Undifferentiated Neuro2a cells were transfected with a pCAG-GFP construct. All groups were counterstained for the DNA marker Hoechst 334. Scale bars: 5 μM.
Fig 3: Confocal imaging of Hsd17b7 subcellular localization in undifferentiated Neuro2a cells. (A–D) Comparison of Hsd17b7-GFP and pCAG-GFP localization with the nuclear marker Emerin (A,B) and Golgi marker RCAS1 (C,D) in fixed cells. DNA counterstain was done with Hoechst 3342. (E,F) Comparison of Hsd17b7-GFP and pCAG-GFP localization in live cells using the lysosomal marker Lysoview. Scale bars: 5 μM.
Supplier Page from Sino Biological, Inc. for Mouse HSD17B7 Gene ORF cDNA clone expression plasmid, N-GFPSpark tag