plc β  (Bioss)

Journal: Frontiers in Neuroanatomy

Article Title: Up-regulation of CB 1 cannabinoid receptors located at glutamatergic terminals in the medial prefrontal cortex of the obese Zucker rat

doi: 10.3389/fnana.2022.1004702

Figure Lengend Snippet: Primary antibodies used.

Article Snippet: CB 1 cannabinoid receptor, G i/o α-subunits, and phospholipase C-β 1 (PLC-β 1 ) were detected by immunoblotting with the following antibodies: goat polyclonal anti-CB 1 receptor (CB1-Go-Af450; Frontier Science Co. Ltd, Hokkaido, Japan), rabbit polyclonal anti-Gα o (sc-387; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-Gα i 1 (sc-391; Santa Cruz Biotechnology), rabbit polyclonal anti-Gα i 2 (sc-7276; Santa Cruz Biotechnology), rabbit polyclonal anti-Gα i 3 (sc-262; Santa Cruz Biotechnology) and mouse monoclonal anti-PLC-β 1 (BD Transduction Laboratories, San Diego, CA, USA) ( ).

Techniques: Affinity Purification, Transduction

Journal: Biomedicines

Article Title: Klotho Modulates Pro-Fibrotic Activities in Human Atrial Fibroblasts through Inhibition of Phospholipase C Signaling and Suppression of Store-Operated Calcium Entry

doi: 10.3390/biomedicines10071574

Figure Lengend Snippet: Effects of Klotho (100 pM) and selective TRPC6 inhibitor (BI-749327, 1 μM) on ionic currents of transient receptor potential canonical channel (I TRPC ) in human atrial fibroblasts. ( A ) The current tracings and I–V relationship of I TRP showed that BI-749327-treated fibroblasts (1 μM, 48 h, n = 13–15 from 4 independent experiments), Klotho-treated fibroblasts (100 pM, 48 h, n = 15 from 3 independent experiments), and combined Klotho and BI-749327 ( n = 15 from 3 independent experiments) had similarly lower peak inward currents and similarly lower peak outward currents compared with the control ( n = 13 from 4 independent experiments). ( B ) Western blots showed a similar expression of TRPC6 and total phospholipase C (PLC) between fibroblasts treated with Klotho (100 pM, 48 h) and control fibroblasts. However, the phosphorylated PLC (p-PLC) expression was lower in the fibroblasts treated with Klotho (100 pM, 48 h) ( n = 3 from 3 independent experiments). ( C , D ) Example photographs and average data ( n = 9 from 3 independent experiments) showed that human atrial fibroblasts treated with BI-749327 (1 μM, 48 h), Klotho (100 pM, 48 h), and combined Klotho and BI-749327 exhibited similarly less migratory capability than the control. Arrows indicate migrating fibroblasts in the wound space. GAPDH was used as a loading control. PLCβ3, phospholipase C beta 3; pPLCβ3, phosphorylated phospholipase C beta 3; TRPC6, transient receptor potential canonical channel 6. * p < 0.05, *** p < 0.005.

Article Snippet: We probed all blots with primary antibodies against procollagen type IA1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), procollagen type III (Abcam, Cambridge, UK), α-smooth muscle actin (α-SMA) (Abcam, Cambridge, UK), TRPC6 (Alomone Labs, Jerusalem, Israel), human PLC beta 3 (PLCβ3) (R&D Systems, Minneapolis, MN, USA), phospho-PLC beta 3 (Ser537) (pPLCβ3 Ser 537) (Cell Signaling Technology, Beverly, MA, USA); secondary antibodies were all conjugated with horseradish peroxidase.

Techniques: Control, Western Blot, Expressing

Journal: Biomedicines

Article Title: Klotho Modulates Pro-Fibrotic Activities in Human Atrial Fibroblasts through Inhibition of Phospholipase C Signaling and Suppression of Store-Operated Calcium Entry

doi: 10.3390/biomedicines10071574

Figure Lengend Snippet: Effects of Klotho (100 pM) and selective TRPC6 inhibitor (BI-749327, 1 μM) on ionic currents of transient receptor potential canonical channel (I TRPC ) in human atrial fibroblasts. ( A ) The current tracings and I–V relationship of I TRP showed that BI-749327-treated fibroblasts (1 μM, 48 h, n = 13–15 from 4 independent experiments), Klotho-treated fibroblasts (100 pM, 48 h, n = 15 from 3 independent experiments), and combined Klotho and BI-749327 ( n = 15 from 3 independent experiments) had similarly lower peak inward currents and similarly lower peak outward currents compared with the control ( n = 13 from 4 independent experiments). ( B ) Western blots showed a similar expression of TRPC6 and total phospholipase C (PLC) between fibroblasts treated with Klotho (100 pM, 48 h) and control fibroblasts. However, the phosphorylated PLC (p-PLC) expression was lower in the fibroblasts treated with Klotho (100 pM, 48 h) ( n = 3 from 3 independent experiments). ( C , D ) Example photographs and average data ( n = 9 from 3 independent experiments) showed that human atrial fibroblasts treated with BI-749327 (1 μM, 48 h), Klotho (100 pM, 48 h), and combined Klotho and BI-749327 exhibited similarly less migratory capability than the control. Arrows indicate migrating fibroblasts in the wound space. GAPDH was used as a loading control. PLCβ3, phospholipase C beta 3; pPLCβ3, phosphorylated phospholipase C beta 3; TRPC6, transient receptor potential canonical channel 6. * p < 0.05, *** p < 0.005.

Article Snippet: We probed all blots with primary antibodies against procollagen type IA1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), procollagen type III (Abcam, Cambridge, UK), α-smooth muscle actin (α-SMA) (Abcam, Cambridge, UK), TRPC6 (Alomone Labs, Jerusalem, Israel), human PLC beta 3 (PLCβ3) (R&D Systems, Minneapolis, MN, USA), phospho-PLC beta 3 (Ser537) (pPLCβ3 Ser 537) (Cell Signaling Technology, Beverly, MA, USA); secondary antibodies were all conjugated with horseradish peroxidase.

Techniques: Control, Western Blot, Expressing