Journal: Bioscience Reports

Article Title: 17β-estradiol binding to ERα promotes the progression of prolactinoma through estrogen-response element-induced CaBP-9k up-regulation

doi: 10.1042/BSR20191330

Figure Lengend Snippet: MMQ and GH3 cells were treated with E2 (1 nM) or E2 + ICI182780 (1 μM) and transfected with luc-ERE, then ( A,B ) the luciferase reporter gene assay was carried out to evaluate the effects of E2 and ERs on CaBP-9k transcription (* P <0.05, compared with NC group, # P <0.05, compared with E2 group).

Article Snippet: After 1 h of incubation with 50 μl protein A, the supernatants were incubated with anti-CaBP-9k antibody (No. sc-74462, Santa Cruz) at 4°C overnight.

Techniques: Transfection, Luciferase, Reporter Gene Assay

Journal: Bioscience Reports

Article Title: 17β-estradiol binding to ERα promotes the progression of prolactinoma through estrogen-response element-induced CaBP-9k up-regulation

doi: 10.1042/BSR20191330

Figure Lengend Snippet: ( A,B ) MMQ and GH3 cells were treated with 0, 0.1, 1 and 10 nmol/l E2 for 24 h, then the cells were collected and submitted to (A,B) Western blotting assay to test the protein expression levels of CaBP-9k, ERα and ERβ (* P <0.05). ( C,D ) GH3 and MMQ cells were treated with 300 nM AZD9496 for 1 h prior to 1 nM E2 treatment, then the cells were harvested and subjected to Western blotting to test the protein expressions of CaBP-9k, ERα and ERβ (* ,+ P <0.05, compared with NC group; # P<0.05, compared with E2 group).

Article Snippet: After 1 h of incubation with 50 μl protein A, the supernatants were incubated with anti-CaBP-9k antibody (No. sc-74462, Santa Cruz) at 4°C overnight.

Techniques: Western Blot, Expressing

Journal: Bioscience Reports

Article Title: 17β-estradiol binding to ERα promotes the progression of prolactinoma through estrogen-response element-induced CaBP-9k up-regulation

doi: 10.1042/BSR20191330

Figure Lengend Snippet: ( A ) IP assay was carried out to analyze the interaction between CaBP-9k protein and ERα/ERβ protein in GH3 cells. ( B ) The subcellular location of CaBP-9k and ERα proteins in GH3 cells were determined by immunofluorescence.

Article Snippet: After 1 h of incubation with 50 μl protein A, the supernatants were incubated with anti-CaBP-9k antibody (No. sc-74462, Santa Cruz) at 4°C overnight.

Techniques: Immunofluorescence

Journal: Bioscience Reports

Article Title: 17β-estradiol binding to ERα promotes the progression of prolactinoma through estrogen-response element-induced CaBP-9k up-regulation

doi: 10.1042/BSR20191330

Figure Lengend Snippet: ( A ) The knockdown efficiency of sh-ERα in GH3 and MMQ cells was determined by Western blotting assay. MMQ and GH3 cells were treated with E2, E2+AZD9496, or E2+sh-ERα, then cells were collected for the following assays. ( B,C ) CCK-8 assay used for cell viability detection. ( D,E ) Flow cytometry used for cell apoptosis detection. ( F,H ) Western blotting analysis used for detection of the protein expression of CaBP-9k (* P <0.05, compared with NC group; # P <0.05, compared with E2 group).

Article Snippet: After 1 h of incubation with 50 μl protein A, the supernatants were incubated with anti-CaBP-9k antibody (No. sc-74462, Santa Cruz) at 4°C overnight.

Techniques: Knockdown, Western Blot, CCK-8 Assay, Flow Cytometry, Expressing

Journal: Bioscience Reports

Article Title: 17β-estradiol binding to ERα promotes the progression of prolactinoma through estrogen-response element-induced CaBP-9k up-regulation

doi: 10.1042/BSR20191330

Figure Lengend Snippet: ( A ) The knockdown efficiency of sh-CaBP-9k was determined by Western blotting assay in both MMQ and GH3 cell lines. Then, MMQ and GH3 cells were treated with E2, sh-CaBP-9k or E2+sh-CaBP-9k and then were submitted to the following assays. ( B,C ) CCK-8 assay was used to detect cell viability. ( D,E ) Flow cytometry was performed to detect cell apoptosis (* P <0.05, compared with NC group, # P <0.05, compared with E2 group).

Article Snippet: After 1 h of incubation with 50 μl protein A, the supernatants were incubated with anti-CaBP-9k antibody (No. sc-74462, Santa Cruz) at 4°C overnight.

Techniques: Knockdown, Western Blot, CCK-8 Assay, Flow Cytometry

Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet: (A–F) Immunohistochemistry technique for capturing SNPH intrusion in PC dendrites. Shown is SNPH (green), Syt2 (red), and Calbindin (blue) labeling in 3.5-month-old WT (A, C, and E) and Shi (B, D, and F) mice. Scale bar, 10 μm. (G and H) High magnification of the maximum intensity projection image from the z stack through dendritic regions of WT (G) and Shi (H). (I and J) Orthogonal (slice) view of SNPH punctum (indicated by arrows in panels G and H) in the dendritic region from WT (I) and Shi (J) in x-z and y-z orientations, respectively. (K) Quantification of percentage area occupied by SNPH within the dendritic volume from 3 mice of each group. Data are shown as mean ± SEM. *p < 0.05. (L–N) Capturing SNPH intrusion by pre-tagging dendritic mitochondria in vivo using viral transduction. (L) Technique to selectively transduce PCs with AAV-Mito-mCherry. (M) Demonstration of successful pre-tagging of mitochondria in dendrites of a single PC by Calbindin staining. Scale bar, 10 μm. (N) Demonstration of how pre-tagged dendritic mitochondria in PCs are used to capture SNPH intrusion by 3D rotation. (O and P) Single dendritic tree in WT (O) or Shi (P) pre-tagged with Mito-mCherry (red) and SNPH intrusion (green) captured by co-rotation with Mito-mCherry. Merged images show the fraction of dendritic mitochondria anchored by intruded SNPH (yellow). (Q and R) Percentage of PCs with SNPH intrusions (Q) and percentage of SNPH bound to mitochondria per dendritic tree (R) from WT (n = 73) and Shi (n = 106) PCs. Data are shown as mean ± SEM. ***p < 0.005.

Article Snippet: Polyclonal Calbindin , Boster , M03047–2.

Techniques: Immunohistochemistry, Labeling, In Vivo, Transduction, Staining

Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet: (A and B) Representative images of lentivirally transduced GFP-SNPH (1–469) (A) and GFP-SNPH (B) in PCs of SNPH-KO mice injected with saline (no harmaline, vehicle only). (C-H) Effect of harmaline on GFP-SNPH (1–469)-transduced (C) and GFP-SNPH-transduced (F) PC dendrites. Degenerating dendrites in GFP-SNPH-transduced PCs can be seen in (F). Also shown is Calbindin labeling of GFP SNPH (1–469) (D) and GFP-SNPH (G) from (C) and (F). Merged images of GFP SNPH (1–469) and GFP-SNPH with Calbindin are shown in (E) and (H), respectively. (I–K) Representative image of a harmaline-induced degenerating PC (white arrow in I) transduced with GFP-SNPH. Calbindin staining from the same section is shown in (J), whereas a merged image is shown in (K). Scale bars, 20 μm. (L) Quantification of dendritic shrinkage in GFP-SNPH (1–469)- and GFP-SNPH-transduced PCs in the absence (n = 3 mice, vehicle only) or presence of harmaline (n = 5 mice). Data are shown as mean ± SEM. ***p < 0.001.

Article Snippet: Polyclonal Calbindin , Boster , M03047–2.

Techniques: Injection, Saline, Labeling, Transduction, Staining

Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet:

Article Snippet: Polyclonal Calbindin , Boster , M03047–2.

Techniques: Virus, Plasmid Preparation, Recombinant, Software, Imaging

Journal: PLoS Genetics

Article Title: Rgs6 is Required for Adult Maintenance of Dopaminergic Neurons in the Ventral Substantia Nigra

doi: 10.1371/journal.pgen.1004863

Figure Lengend Snippet: (A) Rgs6 expression revealed by immunohistofluorescence (red) together with TH (green) in mDA neurons of SNc but not VTA. Scale bar 410 µm. (B) Co-immunofluorescence analysis of TH (green) and Rgs6 or Pitx3 (red, as indicated) in SNc and VTA of adult mice. Arrowheads point to Pitx3-negative or Rgs6-negative mDA neurons in dorsal SNc. Scale bar 100 µm. (C) Triple immunofluorescence staining for TH (green), Pitx3 (red) and Calb1 (blue) on tissue sections of adult mice indicates that the majority of TH+Pitx3− cells of dSNc (arrowheads) are positive for Calb1, while TH+Pitx3+ cells of vSNc are negative for Calb1, consistent with depiction in A. Scale bar 100 µm.

Article Snippet: Primary antibodies used were against Pitx3 (rabbit home-made, 1∶400), Rgs6 (rabbit home-made 1∶25), Th (Millipore MAB318, 1∶1000), Th (Millipore AB152, 1∶500), Calb1 (R&D Systems AF3320, 1∶40), Fgf10 (Millipore ABN44, 1∶1000), Slc6a3 (Santa Cruz sc-32258, 1∶250), Drd2 (Santa Cruz sc5303, 1∶100), Aldh1a1 (Abcam ab24343, 1∶400), BDNF (Abcam ab108319, 1∶25), phospho-p27 (Abcam ab32096, 1∶50), phospho-Erk1/2 (Cell Signaling 4376, 1∶25), DJ-1 (Abcam ab18257, 1∶500) Lrrk2 (Epitomics 3514-1, 1∶50), Pink1 (Novus Biologicals BC100-494, 1∶50), LC3B (Cell Signaling 3868, 1∶50), and Vmat2 (Millipore AB1598P, 1∶200).

Techniques: Expressing, Immunohistofluorescence, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: The temporal progression of retinal degeneration and early-stage idebenone treatment in the Pde6b rd1/rd1 mouse model of retinal dystrophy

doi: 10.1038/s41598-024-52391-y

Figure Lengend Snippet: Antibodies utilized for immunohistochemical and immunofluorescent staining.

Article Snippet: Calbindin D28K , Novus , 1:1000/1:1000 , Chicken , NBP2-50028.

Techniques: Immunohistochemical staining, Staining