pan  (Alomone Labs)

Journal: Nature Communications

Article Title: CYFIP1 governs the development of cortical axons by modulating calcium availability

doi: 10.1038/s41467-025-65801-0

Figure Lengend Snippet: a CYFIP1 RNA immunoprecipitation (RNA-IP) from DIV 3 WT cortical neurons. Histogram showing relative enrichment of the mRNAs over the non-specific IgG, measured by RT-qPCR of the eluate. The values were normalized for the input and mHprt1 mRNA and expressed as fold change over the non-specific IgG of each mRNA ( n = 4 embryos; mean ± SEM; One-Way ANOVA p < 0.0001; mMap1b mRNA p = 0.0390, mCacna1c mRNA p = 0.0054, mCacna1e mRNA p = 0.0078, mCacna1i mRNA p = 0.0009, mCacng2 mRNA p = 0.9997, mCacnb3 mRNA p = 0.9983). b Total mRNA levels of the Ca 2+ channels in DIV 3 WT and Cyfip1 +/- cortical neurons. Histograms represent mCacna1c , mCacna1e , mCacna1i , mCacng2, mCacnb3 and mCyfip1 mRNA levels, normalized to mH3f3 levels and expressed as a fold change over WT (WT n = 6/7 embryos, Cyfip1 +/- n = 7 embryos; mean ± SEM; Two-tailed Multiple Mann-Whitney test, mCacna1c mRNA p = 0.0766, mCacna1e mRNA p = 0.0435, mCacna1i mRNA p = 0.0202, mCacng2 mRNA p = 0.6282, mCacnb3 mRNA p = 0.5343, mCyfip1 mRNA p = 0.0034). c Left, representative Western Blot showing CYFIP1, Ca V 1.2 (CACNA1C), Ca V 2.3 (CACNA1E), Ca V 3.3 (CACNA1I), Ca V γ2 (CACNG2/Stargazin) and Ca V β3 (CACNB3) in membrane-enriched fractions from WT and Cyfip1 +/- DIV 3 cortical neurons. The molecular weight of each protein is indicated in kDa. Right, histogram representing Ca V 1.2, Ca V 2.3, Ca V 3.3, Ca V γ2, Ca V β3 and CYFIP1 protein expression levels in membrane-enriched fractions from WT and Cyfip1 +/- DIV 3 cortical neurons. Protein levels were normalized to Coomassie staining (WT n = 4 embryos, Cyfip1 +/- n = 7/8 embryos; mean ± SEM; Two-tailed Multiple unpaired t -test, Ca V 1.2 p = 0.0338, Ca V 2.3 p = 0.0281, Ca V 3.3 p = 0.0129, Ca V γ2 p = 0.2574, Ca V β3 p = 0.6259, CYFIP1 p = 0.0137). d–f Representative images from WT and Cyfip1 +/- DIV 3 cortical neurons stained for Ca V 1.2, Ca V 2.3, Ca V 3.3 (magenta) and βIII-Tubulin (green) (scale bar 20 μm). Histograms show the fluorescence intensity of each calcium channel normalized to βIII-Tubulin in the total neuron (left) and in the axon (right), expressed as a percentage over WT (Ca V 1.2: WT n = 4 embryos, Cyfip1 +/- n = 5 embryos; mean ± SEM; Two-tailed Mann-Whitney test, total p = 0.1111, axon p = 0.4127; Ca V 2.3: WT n = 5 embryos, Cyfip1 +/- n = 4 embryos; mean ± SEM; Two-tailed Mann-Whitney test, total p = 0.0635, axon p = 0.0159; Ca V 3.3: WT n = 4 embryos, Cyfip1 +/- n = 4 embryos; mean ± SEM; Two-tailed Mann-Whitney test, total p = 0.0286, axon p = 0.0286). Source data are provided as a Source Data file.

Article Snippet: Membranes were incubated with the following antibodies rabbit anti-CYFIP1 (1:1000; Sigma-Aldrich, #AB6046), rabbit anti-Ca V 1.2 (CACNA1C) (1:500, Alomone Labs, #ACC003), rabbit anti-Ca V 2.3 (CACNA1E) (1:500, Alomone Labs, #ACC006), rabbit anti-Ca V 3.3 (CACNA1I) (1:500, Alomone Labs, #ACC009), rabbit anti-Stargazin (Ca V γ2/CACNG2) (1:500, Alomone Labs, #ACC012) and rabbit anti-Ca V β3 (CACNB3) (1:500, Alomone Labs, #ACC008), HuD/Elavl4 (1:500, Santa Cruz, #sc-48421) and HuR/Elavl1 (1:500, Santa Cruz, #sc-5261).

Techniques: RNA Immunoprecipitation, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY, Western Blot, Membrane, Molecular Weight, Expressing, Staining, Fluorescence

Journal: Nature Communications

Article Title: CYFIP1 governs the development of cortical axons by modulating calcium availability

doi: 10.1038/s41467-025-65801-0

Figure Lengend Snippet: CYFIP1, potentially interacting with the RNA binding proteins HuD and HuR (previously identified as CYFIP1 interactors), is implicated in regulating the mRNA stability of calcium channel subunits Cacna1c , Cacna1e and Cacna1i . In Cyfip1 +/- neurons, reduced CYFIP1 levels result in a decrease protein abundance of the regulated calcium channel subunits, consequently leading to a decrease in intracellular and mitochondrial calcium concentration. Low levels of calcium ions may affect mitochondria polarity and motility, both of which we found impaired in Cyfip1 +/- axons. The decreased calcium concentration and the mitochondrial defects concur in reducing axonal growth observed in Cyfip1 +/- neurons. By restoring the intracellular calcium homeostasis, both the axonal growth and mitochondrial defects are rescued. Created in BioRender. https://BioRender.com/xquv8cy .

Article Snippet: Membranes were incubated with the following antibodies rabbit anti-CYFIP1 (1:1000; Sigma-Aldrich, #AB6046), rabbit anti-Ca V 1.2 (CACNA1C) (1:500, Alomone Labs, #ACC003), rabbit anti-Ca V 2.3 (CACNA1E) (1:500, Alomone Labs, #ACC006), rabbit anti-Ca V 3.3 (CACNA1I) (1:500, Alomone Labs, #ACC009), rabbit anti-Stargazin (Ca V γ2/CACNG2) (1:500, Alomone Labs, #ACC012) and rabbit anti-Ca V β3 (CACNB3) (1:500, Alomone Labs, #ACC008), HuD/Elavl4 (1:500, Santa Cruz, #sc-48421) and HuR/Elavl1 (1:500, Santa Cruz, #sc-5261).

Techniques: RNA Binding Assay, Quantitative Proteomics, Concentration Assay

Journal: PeerJ

Article Title: Role of the transient receptor potential melastatin 4 in inhibition effect of arsenic trioxide on the tumor biological features of colorectal cancer cell

doi: 10.7717/peerj.17559

Figure Lengend Snippet: (A-B) Representative images of western blot and quantification of TRPM4 in ATO treated HCT116 cells after 9-Ph stimulation. (C) Quantitative analysis of cell viability in ATO treated HCT116 cells after 9-Ph (0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4 μ M) stimulation. (D) Quantitative analysis of cell viability after treatment with 9-Ph at different concentrations. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs Ctl group, # P < 0.05, ### P < 0.001 vs ATO group; n = 3. Data are represented as the mean ± SD. One-way ANOVA followed by Tukey’s test for post-hoc comparisons was used. n, number of independent cells.

Article Snippet: After blocking with 5% skim milk for 2 h, proteins were detected using rabbit TRPM4 antibody (1:200; ACC-004, Alomone Labs, Jerusalem, Israel) and mouse GAPDH antibody (1:1000; Prosun, China), and blots were detected using an Odyssey infrared imaging system (ODY-3149; LI-COR, Lincoln, NE, USA).

Techniques: Western Blot