Journal: bioRxiv

Article Title: INPP5E interactome reveals novel connections to growth factor signaling

doi: 10.64898/2026.02.13.705725

Figure Lengend Snippet: (a) Lysates from HEK293T cells transiently expressing the indicated proteins (top) were subjected to immunoprecipitation with anti-EGFP beads (IP: EGFP). Immunoprecipitates (IPs) and lysates were then analyzed by Western blot (WB) with antibodies against phosphotyrosine (pY), EGFP, Flag or active SRC (SRC pY419). (b) Similar experiment as in (a), except INPP5E was expressed with a Flag epitope and immunoprecipitated with anti-Flag beads (IP: Flag), while the other proteins were expressed as EGFP fusions. Molecular weight markers are shown on the right (kDa). Note how pY signal in (b) matches size of Flag-INPP5E (≈73 kDa), whereas in (a) it matches that of EGFP-INPP5E (≈100 kDa).

Article Snippet: Mouse anti-alpha-tubulin (Proteintech, 66031-1-Ig), mouse anti-EGFP (Proteintech, 66002-1-Ig), rabbit anti-EGFP (Proteintech, 50430-2-AP), mouse anti-Flag (Proteintech, 66008-3-Ig, or Sigma, F3165 or F1804), rabbit anti-Myc (Proteintech, 16286-1-AP) and mouse anti-V5 (Thermofisher, MA5-15253) were used as described [ , ].

Techniques: Expressing, Immunoprecipitation, Western Blot, FLAG-tag, Molecular Weight

Journal: bioRxiv

Article Title: INPP5E interactome reveals novel connections to growth factor signaling

doi: 10.64898/2026.02.13.705725

Figure Lengend Snippet: (a) PI(4,5)P 2 phosphatase activity assay was performed on anti-EGFP immunoprecipitates from HEK293T cell lysates cotransfected with EGFP-INPP5E and PDGFRα-Flag (WT or D842V), as indicated. EGFP-INPP5E WT and D477N (catalytically inactive mutant) were used as positive and negative controls, respectively. Data are mean±SD of n=3 technical replicates from a single experiment. (b) Same as in (a) but using PIP 3 as substrate for the phosphatase activity assay. The INPP5E-catalyzed 5-phosphatase reactions are shown above for both substrates.

Article Snippet: Mouse anti-alpha-tubulin (Proteintech, 66031-1-Ig), mouse anti-EGFP (Proteintech, 66002-1-Ig), rabbit anti-EGFP (Proteintech, 50430-2-AP), mouse anti-Flag (Proteintech, 66008-3-Ig, or Sigma, F3165 or F1804), rabbit anti-Myc (Proteintech, 16286-1-AP) and mouse anti-V5 (Thermofisher, MA5-15253) were used as described [ , ].

Techniques: Phosphatase Assay, Mutagenesis

Journal: bioRxiv

Article Title: INPP5E interactome reveals novel connections to growth factor signaling

doi: 10.64898/2026.02.13.705725

Figure Lengend Snippet: (a) Lysates from HEK293T cells transiently expressing the indicated EGFP fusion proteins (top) and treated or not with orthovanadate, a tyrosine phosphatase inhibitor, were immunoprecipitated with anti-EGFP beads (IP: EGFP) and analyzed by Western blot (WB) with anti-phosphotyrosine (pY) and anti-EGFP antibodies, as indicated. Molecular weight markers on the right (kDa).

Article Snippet: Mouse anti-alpha-tubulin (Proteintech, 66031-1-Ig), mouse anti-EGFP (Proteintech, 66002-1-Ig), rabbit anti-EGFP (Proteintech, 50430-2-AP), mouse anti-Flag (Proteintech, 66008-3-Ig, or Sigma, F3165 or F1804), rabbit anti-Myc (Proteintech, 16286-1-AP) and mouse anti-V5 (Thermofisher, MA5-15253) were used as described [ , ].

Techniques: Expressing, Immunoprecipitation, Western Blot, Molecular Weight

Journal: Research

Article Title: Endocannabinoids Block Headache and Anxiety Comorbidity via Two-Pronged Anterior Insular Projections

doi: 10.34133/research.1031

Figure Lengend Snippet: Knocking down MAGL in vAI-PrL and dAI-ovBNST synapses ameliorates headache and comorbid anxiety, respectively. (A to C and N to P) Experimental schedule for MAGL knockdown in vAI-PrL (A and B) and dAI-ovBNST (N and O) synapses, viral injections, and behavioral tests. Representative images show mCherry-expressing neurons in the AI (C and P) and EGFP-expressing neurons in PrL (C) and ovBNST (P). Scale bars, 500 μm. (D to F and Q to S) RNAscope and immunofluorescence costaining show the effective knockdown of the expression of MAGL mRNA in vAI-PrL (D to F) and dAI-ovBNST (Q to S) circuits. Dotted coils indicate neurons infected with AAV viruses. Scale bars, 100 μm (top) or 20 μm (bottom). (G and T) Von Frey tests show effects of MAGL-KD in vAI-PrL (G) or dAI-ovBNST (T) on cephalic cutaneous allodynia. (H to M and U to Z) OFT (H to J and U to W) and EPM tests (K to M and X to Z) show effects of MAGL-KD in vAI-PrL (H to M) or dAI-ovBNST (U to Z) on anxiety-like behaviors. MAGL-NC, MAGL noncoding control; MAGL-KD, MAGL knockdown. The data are presented as the mean ± SEM. ** P <0.01 versus MAGL-NC (F and S). * P <0.05, ** P <0.01 versus Vehicle + MAGL-NC (G to M and T to Z). Detailed statistical results are provided in Table .

Article Snippet: The following antibodies for immunofluorescence staining were employed: rabbit anti-c-fos primary antibodies (1:3,000, ab279289, Abcam), rabbit EGFP [1:100, 2555, Cell Signaling Technology (CST)] primary antibodies (1:100, 43590, CST), rabbit anti-mCherry primary antibodies (1:100, 43590, CST), donkey anti-rabbit Alexa Fluor 488 secondary antibodies (1:1,000, ab150113, Abcam), and donkey anti-rabbit Alexa Fluor 568 secondary antibodies (1:1,000, ab175470, Abcam).

Techniques: Knockdown, Expressing, RNAscope, Immunofluorescence, Infection, Control