Journal: Neuroscience Bulletin

Article Title: Quantification of Tyrosine Hydroxylase and ErbB4 in the Locus Coeruleus of Mood Disorder Patients Using a Multispectral Method to Prevent Interference with Immunocytochemical Signals by Neuromelanin

doi: 10.1007/s12264-019-00339-y

Figure Lengend Snippet: Flowchart of the multispectral method used to prevent the interference of neuromelanin with the immunocytochemical signal of tyrosine hydroxylase (TH) or ErbB4 in postmortem human locus coeruleus (LC) tissue. MDD, major depressive disorder; BD, bipolar disorder; CNTR, control group; IOD, integrated optical density.

Article Snippet: The next day, the sections were rinsed in TBS for 3 × 10 min and subsequently incubated with anti-rabbit IgG conjugated to alkaline phosphatase (1:200) at RT for 1 h. ErbB4 was stained blue using a Fast Blue kit (cat. No. SK-5300, Vector Laboratories, Burlingame, CA) for 50 min. Then the sections were rinsed for 3 × 5 min and incubated with biotinylated anti-mouse (1:200) at RT for 1 h and with avidin-biotin complexes (ABC, 1:800) at RT for 1 h. TH was stained red using the 3-amino-9-ethylcarbazole (AEC) red kit (cat. No. SK-4200, Vector Laboratories) for 50 min.

Techniques:

Journal: Neuroscience Bulletin

Article Title: Quantification of Tyrosine Hydroxylase and ErbB4 in the Locus Coeruleus of Mood Disorder Patients Using a Multispectral Method to Prevent Interference with Immunocytochemical Signals by Neuromelanin

doi: 10.1007/s12264-019-00339-y

Figure Lengend Snippet: ErbB4 immunoreactivity (ir) in the locus coeruleus (LC) neurons of mood disorder patients, and co-localization of tyrosine hydroxylase (TH)-ir and ErbB4-ir in the human LC. A ErbB4-ir (blue) was present in the cytoplasm in neurons containing neuromelanin (dark brown) both in controls and mood disorder patients. The distribution and appearance of the ErbB4-ir neurons in mood disorder patients were similar to those of controls. B There was no difference between the IODs of ErbB4-ir in MDD patients and their controls, or BD patients and their controls. C Original microscope image showing signals of TH-ir (red), ErbB4-ir (blue), and neuromelanin (brown). D After color separation, the TH-ir (red) and ErbB4-ir (blue) signals are presented in one image. Co-localization (yellow) was observed in a considerable number of LC neurons. The data are shown as mean + SD. IOD, integrated optical density; CTR1, control group for BD; CTR2, control group for MDD; scale bar, 200 μm.

Article Snippet: The next day, the sections were rinsed in TBS for 3 × 10 min and subsequently incubated with anti-rabbit IgG conjugated to alkaline phosphatase (1:200) at RT for 1 h. ErbB4 was stained blue using a Fast Blue kit (cat. No. SK-5300, Vector Laboratories, Burlingame, CA) for 50 min. Then the sections were rinsed for 3 × 5 min and incubated with biotinylated anti-mouse (1:200) at RT for 1 h and with avidin-biotin complexes (ABC, 1:800) at RT for 1 h. TH was stained red using the 3-amino-9-ethylcarbazole (AEC) red kit (cat. No. SK-4200, Vector Laboratories) for 50 min.

Techniques: Microscopy

Journal: Frontiers in Pharmacology

Article Title: Synergistic Activity of the HSP90 Inhibitor Ganetespib With Lapatinib Reverses Acquired Lapatinib Resistance in HER2-Positive Breast Cancer Cells

doi: 10.3389/fphar.2021.651516

Figure Lengend Snippet: Synergistic effect of ganetespib and lapatinib in lapatinib-sensitive and -resistant breast cancer cell lines. (A) SKBR3, BT474, (B) SKBR3-L, and BT474-L cells were treated with ganetespib, lapatinib or ganetespib plus lapatinib at the indicated optimal combinative concentrations obtained from the Chou-Talalay method (CompuSyn software). Data are presented as means ± SD of three independent experiments, * p < 0.05, ** p < 0.01 vs . combination administration for each cell line by ANOVA. (C) SKBR3, SKBR3-L, (D) BT474, and BT474-L cells were incubated with ganetespib, lapatinib or combination for 24 h. Protein levels were analyzed by western blot for EGFR, HER2, HER3, Akt, ERK and their phosphorylated forms.

Article Snippet: The HER/ErbB family Antibody Sample Kit from Cell Signaling Technology included antibodies against EGFR (D38B1), HER2/ErbB2 (D8F12), HER3/ErbB3 (D22C5), HER4/ErbB4 (111B2), Phospho-EGFR (Tyr1068), Phospho-HER2/ErB2 (Tyr1221/1222), Phospho-HER3/ErB3 (Tyr1289), and Phosphor-HER4/ErbB4 (Tyr1284).

Techniques: Software, Incubation, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Synergistic Activity of the HSP90 Inhibitor Ganetespib With Lapatinib Reverses Acquired Lapatinib Resistance in HER2-Positive Breast Cancer Cells

doi: 10.3389/fphar.2021.651516

Figure Lengend Snippet: Antitumor activity of ganetespib and lapatinib, alone or in combination, in xenograft models in vivo . (A) and (B) Bearing tumors with volumes of approximately 50 mm3, SKBR3 and SKBR3-L tumor xenograft nude mice were randomized into treatment groups (n = 6/group) and received vehicle, lapatinib, ganetespib or combination (in a vehicle of 1% DMSO, 30% polyethylene glycol and 1% Tween 80) for 4 weeks. Data are presented as means ± SD (n = 6, p < 0.05). (C) and (D) Mouse body weight was measured every three days. HER2 expression in (E) SKBR3 and (F) SKBR3-L tumors was determined by immunohistochemistry. Pictures a and e show vehicle controls; pictures b and f, c and g, d and h show lapatinib-, ganetespib-, and combination-treated groups for the SKBR3 and SKBR3-L xenografts, respectively. Tumor tissues excised from (G) SKBR3 and (H) SKBR3-L xenografts were lyzed, and changes in the levels of EGFR, HER2, HER3, Akt, ERK, and STAT3 protein were assessed by western blot.

Article Snippet: The HER/ErbB family Antibody Sample Kit from Cell Signaling Technology included antibodies against EGFR (D38B1), HER2/ErbB2 (D8F12), HER3/ErbB3 (D22C5), HER4/ErbB4 (111B2), Phospho-EGFR (Tyr1068), Phospho-HER2/ErB2 (Tyr1221/1222), Phospho-HER3/ErB3 (Tyr1289), and Phosphor-HER4/ErbB4 (Tyr1284).

Techniques: Activity Assay, In Vivo, Expressing, Immunohistochemistry, Western Blot