anti p plcγ 1  (Cell Signaling Technology Inc)

Journal: Cell Death & Disease

Article Title: Melanophilin-induced primary cilia promote pancreatic cancer metastasis

doi: 10.1038/s41419-025-07344-2

Figure Lengend Snippet: A , B PLCG1 was activated in -QQ PANC-1 cells. A Phosphorylated PLCG1 increased as shown by human phospho-kinase array analysis in Prt and -QQ PANC-1 cells. The red line shows reference spots, whereas the blue line indicates the phosphorylation of Y783 of PLCG1. B PLCG1 was phosphorylated at tyrosine 783 in -QQ cells. Extracts of parental (Prt) or -QQ PANC-1 cells were analyzed by western blot assay with antibodies against phosphorylated PLCG1 at Y783, PLCG1, and tubulin. C The PLCG1 mRNA level increased in -QQ cells. Quantitative results of relative phospholipase C family mRNA levels were analyzed by qPCR in Prt or -QQ PANC-1 cells. D – F PLCG1 promoted EMT, cell migration, and invasion. D Depletion of PLCG1 alleviated EMT in -QQ cells. Extracts of -QQ PANC-1 cells transfected with siRNA against PLCG1 (siPLCG1) were analyzed by western blot assay with antibodies against PLCG1, E-cadherin (E-cad), and Ku70. E , F Depletion of PLCG1 alleviated cell migration and invasion of -QQ cells. Quantitative results of the relative ( E ) migrated and ( F ) invaded cell numbers in Prt or -QQ PANC-1 cells in the presence or absence of siRNA against PLCG1 (siPLCG1). G Depletion of IFT88 decreased PLCG1 expression. Extracts of -QQ PANC-1 cells in the absence or presence of siRNA against IFT88 (siIFT88) were analyzed by western blot assay with antibodies against phosphorylated PLCG1 (Y783), PLCG1, and Ku70. H MLPH depletion decreased PLCG1 expression and restored EMT. Extracts of -QQ-shScr or -QQ-shMLPH#1 or #2 PANC-1 cells were analyzed by western blot assay with antibodies against MLPH, PLCG1, E-cadherin (E-cad), and actin. I – J PLCG1 and phosphorylated PLCG1 at Y783 (p-PLCG1) localized to the primary cilia. Immunofluorescence staining of Prt or -QQ PANC-1 cells with antibodies against acetylated tubulin (Ac-tub) and ( I ) p-PLCG1 and ( J ) PLCG1. DNA was stained with DAPI (blue). Scale bar, 10 µm. K Depletion of PLCG1 inhibited primary ciliogenesis. Quantitative results of the frequency of ciliated cells of Prt or -QQ PANC-1 cells were shown in the presence or absence of siRNA against PLCG1 (siPLCG1). L Graphic abstract. When PDAC cells suffered a Gln deprivation condition, MLPH was upregulated and recruited to the centrosome to facilitate primary ciliogenesis. The primary cilia induced and activated PLCG1 to promote EMT, invasion, and metastasis. Besides, PLCG1 was recruited to the primary cilia, thereby feedforward maintaining primary ciliogenesis. Data are represented as the mean ± SD of three independent experiments. n.s. no significance, * P < 0.05, *** P < 0.001.

Article Snippet: Anti-IFT88 (13967-1-AP), anti-ARL13B (17711-1-AP), anti-MLPH (10338-1-AP), and anti-MMP9 (10375-2-AP) antibodies were purchased from Proteintech (Chicago, IL, USA). anti-CEP164 (NBP1-81445) and anti-myosin 5a (NBP1-92156) antibodies were purchased from Novus (Littleton, CO, USA). anti-β-actin (AC-15; GTX26276) and anti-PLCγ-1 (phospho-Y783; GTX24828) were purchased from GeneTex (Irvine, CA, USA).

Techniques: Western Blot, Migration, Transfection, Expressing, Immunofluorescence, Staining

Journal: Brain research bulletin

Article Title: Role of BDNF-TrkB signaling in the improvement of motor function and neuroplasticity after ischemic stroke in rats by transcranial direct current stimulation.

doi: 10.1016/j.brainresbull.2024.111164

Figure Lengend Snippet: Fig. 4. Effects of tDCS application on activation of PLCγ / CaMK IV / CREB and ERK1/2 / RSK signaling cascade following MCAO/R. (A) Plasma BDNF levels of patients. T test was used for statistical analysis. (B) Tricolor fluorescent labeling BDNF, NeuN, and DAPI, to observe the ischaemic semi-dark band in the infarcted region of the left cerebral cortex. Representative Western bands were normalised to the corresponding total protein levels and to GAPDH as a control, (C) Quan tification of the relative protein levels of PLCγ1, CaMK IV and CREB1 in three groups. The original images of the Western blot bands are available in the supple mentary material. (D) PLCγ1 expression,(E) CaMK IV expression, and (F) CREB1 expression in different groups. (G) Representative Western bands were normalised to the corresponding total protein levels and to GAPDH as a control. Quantification of the relative proteins levels of P-ERK, ERK and RSK in three groups. The original images of the Western blot bands are available in the supplementary material.(H) P-ERK1/2 / ERK1/2 expression and (I) RSK expression in different groups. The data are presented as means ± SEM. *p < 0.05,* *p < 0.01 vs. MCAO/R group. Differences between the groups were analyzed using one-way ANOVA followed by Tukey’s post hoc test. Different coloured bars represent different subgroups, and the vertical values show the ratio of the expression of the relevant proteins to the expression of the corresponding internal reference proteins.

Article Snippet: Then membranes were incubated using primary antibodies: anti- PLCγ (1:1000, Proteintech China), anti-CERB (1:1000 Cell Signaling Technology, US), anti-CaMK IV (1:1000, Abcam, UK), anti-RSK (1:1000, Cell Signaling Technology, US), anti-P-ERK1/2 (1:1000, Cell Signaling Technology, US), anti-ERK1/2 (1:1000, Cell Signaling Technology, US), anti-GAP 43 (1:1000, Abcam, UK) and anti-MAP 2 (1:1000, Abcam, UK) at 4◦C overnight.

Techniques: Activation Assay, Clinical Proteomics, Labeling, Western Blot, Control, Expressing