rabbit  (Alomone Labs)

Journal: Scientific Reports

Article Title: Vacuolar-ATPase-mediated muscle acidification caused muscular mechanical nociceptive hypersensitivity after chronic stress in rats, which involved extracellular matrix proteoglycan and ASIC3

doi: 10.1038/s41598-023-39633-1

Figure Lengend Snippet: Changes in RCS-induced decrease of MMWT by antagonists to ASIC3 and TRPs ( A ), in mRNAs expression of ASIC3 and TRPV1 ( B ) and in protein of ASIC3 in DRGs after RCS ( C , D ). ( A ) The presentation methods are the same as those in Fig. . The arrow shows the period at which APETx2 and RR were injected (3–5 d after RCS). Two-way ANOVA with repeated measures (see Supplementary Table ) followed by Bonferroni’s post hoc test. ## : p < 0.01 compared to the PBS group (main effect of the “drug”), $$, $$$: p < 0.01, 0.001 compared to “pre RCS”, *, **: p < 0.05, 0.01 compared to “pre Inj” in each drug group, &&&: p < 0.001 compared to PBS at the time point, APETx2 (APET) 0.73 μM and 2.2 μM reversed the mechanical withdrawal threshold decreased after RCS but RR neither 1.5 μM nor 15 μM did. ( B ) Vertical axis: ASIC3 mRNA and TRPV1 mRNA relative to GAPDH mRNA. DRGs were removed from rats 1 week after RCS. *: p < 0.05. Note that only ASIC3 mRNA level increased significantly after RCS. ( C ) Western blot analysis of ASIC3 protein expression in DRGs of RCS and control (CTR) rats. Blots from two rats for each group were represented. GAPDH was used as an internal control. ( D ) ASIC3 protein levels in DRGs quantified from the Western blot (n = 6 for each group). Data were shown as relative values to GAPDH. * : p < 0.05. (See Supplementary Table for statistics summary for ( B ) and ( D )).

Article Snippet: The upper piece of membrane was probed with anti-ASIC3 antibody raised in rabbit (1:200, GeneTex, Irvine, CA, USA) and goat anti-rabbit IgG conjugated with HRP (1:2000, Sigma Aldrich), and the lower piece with mouse anti-GAPDH antibody (1:10,000, GeneTex) and goat anti-mouse IgG conjugated with HRP (1:1000, Thermo Fisher Scientific Inc.) to use as loading control.

Techniques: Expressing, Injection, Western Blot

Journal: Drug Design, Development and Therapy

Article Title: Zhiqiao Gancao Decoction Ameliorates Hyperalgesia in Lumbar Disc Herniation via the CCL2/CCR2 Signaling Pathway

doi: 10.2147/DDDT.S415127

Figure Lengend Snippet: CCL2 induces the expression of CCR2 and ASIC3 in DRGn, which is inhibited by CCR2 inhibitors and ZQGCD containing serum ( A ) The cell viability of DRGn. After 12 h pretreatment with RS504393 and ZQGCD containing serum (5%, 10%, and 15%), there was no change in cell viability ( P >0.05). After adding CCL2 for 24 h, the viability of unpretreated cells decreased significantly ( P <0.05), while that of pretreated DRGn did not change compared with the control group ( P >0.05). ( B ) Expression of CCR2 and ASIC3 mRNA in DRGn. ( C and D ) Expression of CCR2 and ASIC3 proteins in DRGn. ** P <0.01, *** P <0.001, **** P <0.0001, ns: P >0.05.

Article Snippet: Neutral resin, xylene, methanol, and ethanol (10004160, 1330-20-7, 67-56-1, 64-17-5, Sinoppharmacy Chemical Reagent Co., LTD., Beijing, China), high-temperature resistant plastic dyeing stand, DAB Kit (RAK-2001, DAB-1031, Fuzhou Maixin Biotechnology Development Co., LTD., Fuzhou, China), CCL2, IL-6, TNF-, IL-1 ELISA kit (JEB-13579, LA160102H, LA166601H, LA167616H, Nanjing Lapda Biotechnology Co., LTD., Nanjing, China), SYBR Green I fluorescent dye, AceQ Universal SYBR qPCR Master Mix (Nanjing Nuoweizan Biotechnology Co., LTD., Nanjing, China), RIPA Lysate (P0013C, Shanghai Biyuntian Biotechnology Co., LTD., Shanghai, Shanghai Biyuntian Biotechnology Co., Shanghai Biyuntian Biotechnology Co., Shanghai Biyuntian Biotechnology Co., Shanghai, Shanghai, Shanghai, Shanghai, Shanghai, China), HRP-conjugated Affinipure Goat Anti-Mouse IgG (H+L), -actin Antibody, ASIC3 Antibody (SA00001-2, SA00001-1, 20536-1-AP, A10268 Proteintech Group, Inc., IL, USA).

Techniques: Expressing, Control

Journal: Drug Design, Development and Therapy

Article Title: Zhiqiao Gancao Decoction Ameliorates Hyperalgesia in Lumbar Disc Herniation via the CCL2/CCR2 Signaling Pathway

doi: 10.2147/DDDT.S415127

Figure Lengend Snippet: CCL2/CCR2 pathway and ASIC3 were highly in DRG of LDH rats expressed but inhibited by CCR2 inhibitor or ZQGCD. ( A ) Bar graphs showing the expression levels of CCL2 and CCR2 mRNA in DRG. ( B and C ) Immunoblot and bar graphs showing the expression levels of CCL2 and CCR2 in DRG. * P <0.05, ** P <0.01, **** P <0.0001, ns: P >0.05.

Article Snippet: Neutral resin, xylene, methanol, and ethanol (10004160, 1330-20-7, 67-56-1, 64-17-5, Sinoppharmacy Chemical Reagent Co., LTD., Beijing, China), high-temperature resistant plastic dyeing stand, DAB Kit (RAK-2001, DAB-1031, Fuzhou Maixin Biotechnology Development Co., LTD., Fuzhou, China), CCL2, IL-6, TNF-, IL-1 ELISA kit (JEB-13579, LA160102H, LA166601H, LA167616H, Nanjing Lapda Biotechnology Co., LTD., Nanjing, China), SYBR Green I fluorescent dye, AceQ Universal SYBR qPCR Master Mix (Nanjing Nuoweizan Biotechnology Co., LTD., Nanjing, China), RIPA Lysate (P0013C, Shanghai Biyuntian Biotechnology Co., LTD., Shanghai, Shanghai Biyuntian Biotechnology Co., Shanghai Biyuntian Biotechnology Co., Shanghai Biyuntian Biotechnology Co., Shanghai, Shanghai, Shanghai, Shanghai, Shanghai, China), HRP-conjugated Affinipure Goat Anti-Mouse IgG (H+L), -actin Antibody, ASIC3 Antibody (SA00001-2, SA00001-1, 20536-1-AP, A10268 Proteintech Group, Inc., IL, USA).

Techniques: Expressing, Western Blot