Journal: Pathology International

Article Title: FHL1: A novel diagnostic marker for papillary thyroid carcinoma

doi: 10.1111/pin.13467

Figure Lengend Snippet: Clinical, and histopathological characteristics of patients with papillary thyroid carcinoma (classical subtype).

Article Snippet: Slides were immunostained with monoclonal antibodies against the BCL2 antigen (clone SP66, Maixin), CD117/C‐kit antigen (clone YR145, Maixin), and with monoclonal antibodies against the FHL1 (10991‐1‐AP, Proteintech Group, Inc.), PPARGC1A (Boster, China), and TLE1 antigen (Maixin).

Techniques: Expressing

Journal: Pathology International

Article Title: FHL1: A novel diagnostic marker for papillary thyroid carcinoma

doi: 10.1111/pin.13467

Figure Lengend Snippet: The expression of FHL1, BCL2, TLE1, KIT, PPARGC1A and GHR in papillary thyroid carcinoma (PTC) and the control group. FHL1 showed strong positivity in the nuclei and to a lesser extent in the cytoplasm of normal thyroid tissues adjacent to the tumour (the control group), but is not present in the PTC tissues. ( p < 0.01). There was no significant difference in the expression of BCL2 between PTC and normal thyroid tissues ( p > 0.05). The results showed that TLE1, KIT, PPARGC1A, and GHR were absent in both tumor and normal tissues. (magnification ×200). FHL1, four and a half LIM domains 1.

Article Snippet: Slides were immunostained with monoclonal antibodies against the BCL2 antigen (clone SP66, Maixin), CD117/C‐kit antigen (clone YR145, Maixin), and with monoclonal antibodies against the FHL1 (10991‐1‐AP, Proteintech Group, Inc.), PPARGC1A (Boster, China), and TLE1 antigen (Maixin).

Techniques: Expressing, Control

Journal: Gene

Article Title: Role of HIF-1α in the Hypoxia Inducible Expression of the Thiamine Transporter, SLC19A3

doi: 10.1016/j.gene.2016.10.013

Figure Lengend Snippet: (A) Representative Western blot for HIF-1α, LDHA, THTR1, THTR2 and SP1 expression in BT-474 whole cell lysates after 250µM DFO and 1% O2 exposure for 24, 48, and 72h compared to normoxic (N) control. Actin was used as a loading control. Densitometry of protein expression after (B) DFO or (C) 1% O2 treatment with n=7 independent experiments for HIF-1α and n=3 independent experiments for all other proteins. (⋆) Represents a statistically significant difference of p<0.05 based on the results of a one-way ANOVA with Tukey’s post-hoc test.

Article Snippet: Antibodies used were directed against HIF-1α (Bethyl Laboratories, Montgomery, TX), SP1 (Genetex, Irvine, CA) and a nonspecific IgG (SantaCruz Biotechnology, Dallas, TX).

Techniques: Western Blot, Expressing, Control

Journal: Gene

Article Title: Role of HIF-1α in the Hypoxia Inducible Expression of the Thiamine Transporter, SLC19A3

doi: 10.1016/j.gene.2016.10.013

Figure Lengend Snippet: Transfected BT-474 cells were exposed to 1% O2 for 48h or to normoxia. The dual luciferase assay was used to quantify the level of promoter activity and the results are reported as the fold change +/− standard deviation of the hypoxia to normoxia luciferase activity ratio with n=5 independent experiments. (A) Empty pGL3 Basic and pGL3-HRE as positive control. (B) Full SLC19A2, full SLC19A3 and SLC19A3 deletion constructs. (C) Promoter activity of the full length SLC19A3 mutation promoter constructs. (D) SLC19A3 minimal promoter region. Locations of HREs are indicated by bold and italics while the SP1 binding element is underlined. The location of potential HASs are indicated with a box. The +1 indicates the location of the transcriptional start site. (⋆) Represents a statistically significant difference of p<0.05 based on the results of a Student’s T-test (A) or a one-way ANOVA (B and C) with Tukey’s post-hoc test.

Article Snippet: Antibodies used were directed against HIF-1α (Bethyl Laboratories, Montgomery, TX), SP1 (Genetex, Irvine, CA) and a nonspecific IgG (SantaCruz Biotechnology, Dallas, TX).

Techniques: Transfection, Luciferase, Activity Assay, Standard Deviation, Positive Control, Construct, Mutagenesis, Binding Assay

Journal: Gene

Article Title: Role of HIF-1α in the Hypoxia Inducible Expression of the Thiamine Transporter, SLC19A3

doi: 10.1016/j.gene.2016.10.013

Figure Lengend Snippet: Representative Western blot of HIF-1α and SP1 expression in nuclear lysates after (A) 250µM DFO and (B) 1% O2 exposure for 24, 48, and 72h. P84 was used for loading control. Densitometry of transcription factor protein expression after (C) DFO or (D) 1% O2 exposure up to 72 h. Data are presented with n=5 independent experiments. (⋆) Represents a statistically significant difference of p<0.05 as determined by a one-way ANOVA with Tukey’s post-hoc test. (E) Chromatin Immunoprecipitation assay for BT-474 cells treated with hypoxia (1% O2 or 250µM DFO) for 24h and untreated normoxic controls. Water only reactions containing no chromatin and input reactions were used as negative and positive controls, respectively. HIF-1α and SP1 reactions represent chromatin immunoprecipitated, while a random IgG was used as a negative control. The minimal promoter region of the SLC19A3 promoter was signified by an amplicon of 250bp.

Article Snippet: Antibodies used were directed against HIF-1α (Bethyl Laboratories, Montgomery, TX), SP1 (Genetex, Irvine, CA) and a nonspecific IgG (SantaCruz Biotechnology, Dallas, TX).

Techniques: Western Blot, Expressing, Control, Chromatin Immunoprecipitation, Immunoprecipitation, Negative Control, Amplification

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: SH3GLB1-related autophagy mediates mitochondrial metabolism to acquire resistance against temozolomide in glioblastoma

doi: 10.1186/s13046-022-02429-8

Figure Lengend Snippet: Sp1 promotes SH3GLB1 expression. A Genetic expression of SH3GLB1 and Sp1 from the GBM dataset of the CGGA database is shown in scatter plots with correlation assessment. B Schematic graph suggesting the potential Sp1 binding site on the SH3GLB1 promoter. Enhanced binding of Sp1 to the SH3GLB1 promoter is shown by chromatin immunoprecipitation assay in the resistant cells. C Western blot analysis showing enhanced Sp1 and SH3GLB1 expression in the resistant cells. D Western blot analysis showing reduced SH3GLB1 expression in the resistant cells with siSp1. E Western blot analysis showing that mithramycin A (MA) alleviated the TMZ-induced enhancement of Sp1 and SH3GLB1. F Sorted CD133 + and CD133 − subsets from patient-derived primary GBM cells. The former exhibit higher Sp1, LC3B-II and lower p62 in the Western blot analysis. N = 3 in each group, * p < 0.05

Article Snippet: The detection antibodies were as follows: SH3GLB1 (Proteintech, Rosemont, IL, USA), CD133 (Proteintech), SP1 (Merck Millipore, Burlington, MA, USA), LC3B (Santa Cruz, Dallas, TX, USA), p62 (Cell Signaling, Danvers, MA, USA), caspase 3 (Cell Signaling), actin (Merck Millipore), Atg12 (GeneTex, Irvine, CA, USA), and OXPHOS antibody cocktail (ab110411, Abcam, Cambridge, UK), Vinculin (Thermo Fisher Scientific).

Techniques: Expressing, Binding Assay, Chromatin Immunoprecipitation, Western Blot, Derivative Assay