mcherry primary antibody  (TaKaRa)

Journal: Research

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

doi: 10.34133/research.1031

Figure Lengend Snippet: Chemo-inhibition of PrL- and ovBNST-projecting AI neurons ameliorates ISDN-induced headache and comorbid anxiety, respectively. (A) Schematic diagram for injections of mCherry- and EGFP-expressing retrogradely transported viruses into PrL and ovBNST, respectively. (B) Retrogradely transported viruses labeled PrL- and ovBNST-projecting neurons located in the ventral and dorsal parts of AI, respectively, with bare overlap. Scale bar, 200 μm (left) or 100 μm (right). (C) Statistical analysis shows the percentage of merged neurons in mCherry- or EGFP-expressing AI neurons. (D and M) Experimental schedule and schematic diagram for chemogenetic inhibition of PrL-projecting vAI neurons (D) and ovBNST-projecting dAI neurons (M). (E and N) Representative images show the EGFP-expressing neurons in the PrL (E) or ovBNST (N) and mCherry-expressing neurons in AI. Scale bars, 500 μm (left) or 100 μm (right). (F and O) Von Frey tests show effects of chemogenetic inhibition of PrL-projecting vAI neurons (F) and ovBNST-projecting dAI neurons (O) on cephalic cutaneous allodynia in mice with repeated ISDN injections. (G to L and P to U) OFT (G to I and P to R) and EPM test (J to L and S to U) show effects of chemogenetic inhibition of PrL-projecting vAI neurons (G to L) and ovBNST-projecting dAI neurons (P to U) on anxiety-like behaviors in mice with repeated ISDN injections. The data are presented as the mean ± SEM. * P <0.05, ** P <0.01 versus Vehicle + mCherry. 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: Inhibition, Expressing, Labeling

Journal: Research

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

doi: 10.34133/research.1031

Figure Lengend Snippet: Knockdown of DAGLα in vAI-PrL and dAI-ovBNST synapses has no effect on cephalic cutaneous allodynia and anxiety-like behaviors in ISDN-injected mice. (A to C and N to P) Experimental schedule for DAGLα knockdown in vAI-PrL (A and B) and dAI-ovBNST synapses (N and O), 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 immunofluorescent costaining show the effective knockdown of the expression of DAGLα mRNA in AI-innervating PrL (D to F) and ovBNST (Q to S) neurons. 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 DAGLα-KD in vAI-PrL (G) or dAI-ovBNST (T) on repeated ISDN-induced 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 the effects of DAGLα-KD in vAI-PrL (H to M) or dAI-ovBNST (U to Z) on repeated ISDN-induced anxiety-like behaviors. DAGLα-NC, DAGLα noncoding control; DAGLα-KD, DAGLα knockdown; INs: innervating neurons. The data are presented as the mean ± SEM. ** P <0.01 versus DAGLα-NC (F and S). * P <0.05, ** P <0.01 versus Vehicle + DAGLα-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, Injection, Expressing, RNAscope, Infection, Control

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