phospho irs1 n a n a n a total pten cell signaling technology  (Cell Signaling Technology Inc)

Journal: Experimental Diabetes Research

Article Title: Insulin Receptor Substrate 2 Expression and Involvement in Neuronal Insulin Resistance in Diabetic Neuropathy

doi: 10.1155/2011/212571

Figure Lengend Snippet: IRS isoform expression in murine lumbar DRG. IRS isoforms were examined using RT-PCR ((a) and (b)) and Western blot ((c) and (d)). (a) RT-PCR was performed on adult C57Bl/6 mouse lumbar DRG ( n = 3 mice) and comparisons were made among IRS1, IRS2, IRS3, and IRS4. GAPDH was used as the housekeeping gene. ΔCt values for IRS2 were significantly lower than any other isoform. (b) Analysis of IRS mRNA levels that were normalized to IRS1 mRNA levels revealed that IRS2 mRNA expression was 27-fold higher than IRS1 in the DRG. *denotes P < .05 n = 3 mice. ((c) and (d)) Representative Western blots of IRS1 and IRS2 protein in mouse lumbar DRG. Equal amounts of protein (20 μ g) were loaded for each lane and samples were separated on 4–15% tris-glycine gel. Both IRS1 (c) and IRS2 (d) proteins were readily detectable. All mice were 8-week-old nondiabetic C57Bl/6 males.

Article Snippet: Primary antibodies were used at the following dilutions and incubations: total IRS1 (Santa Cruz, Santa Cruz, CA) 1 : 1000 overnight at 4°C, total IRS2 (Millipore, Billerica, MA) 1 : 500 overnight at 4°C, pSer(731)IRS2 (Abcam, Cambridge, MA) 1 : 1000 overnight at 4°C, insulin receptor β subunit (Santa Cruz) 1 : 500 overnight at 4°C, pSer(473)Akt (Cell Signaling, Danvers, MA) 1 : 500 overnight at 4°C, total Akt (Cell Signaling) 1 : 500 overnight at 4°C, and actin (Millipore) 1 : 100,000 at room temperature for 1 hour.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Nutrition

Article Title: Lonicerae Japonicae Flos extract and chlorogenic acid attenuates high-fat-diet- induced prediabetes via CTRPs-AdipoRs-AMPK/PPARα axes

doi: 10.3389/fnut.2022.1007679

Figure Lengend Snippet: Lonicerae Japonicae Flos and CGA improved the insulin signaling pathway in rat liver. (A–C) Protein levels and densitometric quantification of phospho-insulin receptor substrate 1 (p-IRS1) (Ser307), total(t)-IRS1, phospho-protein kinase B (p-Akt) (Ser473), total(t)-Akt, and β-actin, bars represent densitometric quantification normalized to β-actin. All bars show means ± SEM. ( n = 4 per group). * P < 0.05, vs. CON group; # P < 0.05, vs. HFD group.

Article Snippet: Liver tissues were homogenized in RIPA buffer and centrifuged at 12,000 × g for 20 min. Membranes were incubated with primary antibodies against phospho-IRS1 (Ser307) (p-IRS1) (CST, #2381), total-IRS1 (t-IRS1) (CST, #2382), phospho-Akt (Ser473) (p-Akt) (CST, #4060), total-Akt (t-Akt) (CST, # 4685), AdipoR1 (ab126611), AdipoR2 (ab77612), ELOVL6 (ab69857), AMPK (CST, #5832), p-AMPK (Thr172) (CST, #2535), and peroxisome proliferator-activated receptor-alpha (PPAR-α) (ab24509), and β-actin (C1313).

Techniques: