Journal: Journal of the Endocrine Society

Article Title: Insulin receptor expression and its association with hyperinsulinemia in triple negative breast cancer

doi: 10.1210/jendso/bvag041

Figure Lengend Snippet: Representative images of IHC staining. IHC staining of TNBC specimens was performed as described in the Methods section with antibodies to IR, IGF-1R, pErk1/2, and FOXO3a. The images are shown in 40× magnification. Abbreviations: IGF-1R, IGR-1 receptor; IHC, immunohistochemistry; IR, insulin receptor; pErk1/2, phosphorylated Erk1/2; TNBC, triple negative breast cancer.

Article Snippet: Primary antibodies were anti-IR Ab (ab137747, Abcam, Cambridge, MA, RRID:AB_3717506) 1:300 dilution, anti-IGF-1R Ab (#3027, Cell Signaling, Danvers, CA, RRID:AB_2122378) 1:200 dilution, anti-FOXO3a Ab (#12829, Cell Signaling, RRID:AB_2636990) 1:3200 dilution, and anti-pErk1/2 (Thr202/Tyr204) Ab (#4370, Cell Signaling, RRID:AB_2315112) 1:400 dilution.

Techniques: Immunohistochemistry

Journal: Journal of the Endocrine Society

Article Title: Insulin receptor expression and its association with hyperinsulinemia in triple negative breast cancer

doi: 10.1210/jendso/bvag041

Figure Lengend Snippet: Representative images of protein localization for IR, IGF1R, FOXO3a, and pErk1/2. IHC staining of TNBC specimens was performed and evaluated for localization of proteins. The images are shown in 40× magnification. Abbreviations: IGF-1R, IGR-1 receptor; IHC, immunohistochemistry; IR, insulin receptor; pErk1/2, phosphorylated Erk1/2; TNBC, triple negative breast cancer.

Article Snippet: Primary antibodies were anti-IR Ab (ab137747, Abcam, Cambridge, MA, RRID:AB_3717506) 1:300 dilution, anti-IGF-1R Ab (#3027, Cell Signaling, Danvers, CA, RRID:AB_2122378) 1:200 dilution, anti-FOXO3a Ab (#12829, Cell Signaling, RRID:AB_2636990) 1:3200 dilution, and anti-pErk1/2 (Thr202/Tyr204) Ab (#4370, Cell Signaling, RRID:AB_2315112) 1:400 dilution.

Techniques: Immunohistochemistry

Journal: Journal of the Endocrine Society

Article Title: Insulin receptor expression and its association with hyperinsulinemia in triple negative breast cancer

doi: 10.1210/jendso/bvag041

Figure Lengend Snippet: Overall survival for women with breast cancer based on IR and IGF-1R protein expression in KMPlot.com . KMPlot.com was used to generate overall survival Kaplan-Meier plots. The dataset used was not from our cohort but from Tang and colleagues ; n = 65 patients had IR (A) and IGF1R (B) protein expression available. The patients were split using the “auto select best cutoff” percentile option in KMPlot. No other restrictions were added to the analysis. The plots show hazard ratios with 95% confidence intervals in parentheses . Abbreviations: IGF-1R, IGR-1 receptor; IR, insulin receptor.

Article Snippet: Primary antibodies were anti-IR Ab (ab137747, Abcam, Cambridge, MA, RRID:AB_3717506) 1:300 dilution, anti-IGF-1R Ab (#3027, Cell Signaling, Danvers, CA, RRID:AB_2122378) 1:200 dilution, anti-FOXO3a Ab (#12829, Cell Signaling, RRID:AB_2636990) 1:3200 dilution, and anti-pErk1/2 (Thr202/Tyr204) Ab (#4370, Cell Signaling, RRID:AB_2315112) 1:400 dilution.

Techniques: Expressing

Journal: Biomedicines

Article Title: Insulin and Insulin-like Growth Factor 1 Signaling as a Modulator of MYC Expression in the Meibomian Gland

doi: 10.3390/biomedicines14030578

Figure Lengend Snippet: IIS modulation of MYC and IGF1R in vitro. HMGECs were incubated with trofinetide (100 nM), insulin (2.5 µM), DB1 (5.0 µM), PPP (1.25 µM), BMS (125 nM) and DMSO for 24 h. ( A ) Induction of MYC (Alexa Fluor 488) was demonstrated in HMGECs following incubation with IIS activators (trofinetide, insulin, and DB1) relative to vehicle control (DMSO). Phalloidin: Alexa Fluor 555. ( B ) IGF1R (Alexa Fluor 555) was downregulated in HMGECs following incubation with IIS activators (trofinetide and insulin) relative to vehicle control (DMSO). Phalloidin: Alexa Fluor 488. DAPI (blue). Scale bar: 10 µm.

Article Snippet: Membranes were blocked with 5% BSA in tris-buffered saline (TBS) containing 0.01% tween-20 (TBS-T) for 60 min at RT before incubation with primary antibodies diluted in 1:1000 in 5% BSA in TBS-T overnight at 4 °C with the following antibodies: β-actin (catalog #PA1-183-HRP; ThermoFisher), c-MYC (catalog #5605; Cell Signaling Technology), IGF1Rß (catalog #3027; Cell Signaling Technology), and Akt (catalog #4691 Cell Signaling Technology).

Techniques: In Vitro, Incubation, Control

Journal: Biomedicines

Article Title: Insulin and Insulin-like Growth Factor 1 Signaling as a Modulator of MYC Expression in the Meibomian Gland

doi: 10.3390/biomedicines14030578

Figure Lengend Snippet: IIS modulation of transcript expression in vitro. ( A ) Relative expression of IGF1R was significantly downregulated in trofinetide and DB1-treated HMGECs. Relative ( B ) Akt and ( C ) MYC expression were significantly upregulated compared to DMSO controls (white) following treatment with the IIS activators (light grey) trofinetide and insulin, respectively, while the IIS inhibitors (dark grey) induced significant downregulation of Akt and MYC . * p < 0.05, ** p < 0.01, *** p < 0.001, **** p ≤ 0.0001. 2 −Δ∆cT was utilized to normalize target transcript expression to polR2α.

Article Snippet: Membranes were blocked with 5% BSA in tris-buffered saline (TBS) containing 0.01% tween-20 (TBS-T) for 60 min at RT before incubation with primary antibodies diluted in 1:1000 in 5% BSA in TBS-T overnight at 4 °C with the following antibodies: β-actin (catalog #PA1-183-HRP; ThermoFisher), c-MYC (catalog #5605; Cell Signaling Technology), IGF1Rß (catalog #3027; Cell Signaling Technology), and Akt (catalog #4691 Cell Signaling Technology).

Techniques: Expressing, In Vitro

Journal: Biomedicines

Article Title: Insulin and Insulin-like Growth Factor 1 Signaling as a Modulator of MYC Expression in the Meibomian Gland

doi: 10.3390/biomedicines14030578

Figure Lengend Snippet: Differential transcript expression in the IIS-modulated murine Meibomian gland. Adult mice ( n = 6) were topically treated with trofinetide, PPP, or vehicle, and eyelids were homogenized. There were no significant differences in relative ( A ) IGF1R or ( B ) Akt expression between treatment groups. ( C ) MYC expression was upregulated in trofinetide-treated murine MGs and downregulated in PPP-treated animals relative to respective contralateral vehicle-treated controls. ** p ≤ 0.01. 2 −Δ∆cT was utilized to normalize target transcript expression to polR2α.

Article Snippet: Membranes were blocked with 5% BSA in tris-buffered saline (TBS) containing 0.01% tween-20 (TBS-T) for 60 min at RT before incubation with primary antibodies diluted in 1:1000 in 5% BSA in TBS-T overnight at 4 °C with the following antibodies: β-actin (catalog #PA1-183-HRP; ThermoFisher), c-MYC (catalog #5605; Cell Signaling Technology), IGF1Rß (catalog #3027; Cell Signaling Technology), and Akt (catalog #4691 Cell Signaling Technology).

Techniques: Expressing