Review



manf elisa kit  (Biomatik)


Bioz Verified Symbol Biomatik is a verified supplier
Bioz Manufacturer Symbol Biomatik manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 90
      Buy from Supplier

    Structured Review

    Biomatik manf elisa kit
    (A) Western blot shows the expression of <t>MANF</t> in the normal and injured (3-day transection injury) adult rat DRGs. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows the induction of MANF in injured DRGs (data presented as mean ± SE; standard ‘t’ test; n=3). (C) Co-immunostaining of IB4 (yellow arrows), NF200 (white arrows) and MANF (red arrows) in normal and injured DRGs shows the preferential expression of MANF in IB4 positive non-peptidergic sensory neurons (yellow arrows in the merged view) (scale bar, 50µm).
    Manf Elisa Kit, supplied by Biomatik, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/bio_rxiv__2025__03__14__642904-110-7-14?v=Biomatik
    Average 90 stars, based on 1 article reviews
    manf elisa kit - by Bioz Stars, 2026-07
    90/100 stars

    Images

    1) Product Images from "Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration"

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    Journal: bioRxiv

    doi: 10.1101/2025.03.14.642904

    (A) Western blot shows the expression of MANF in the normal and injured (3-day transection injury) adult rat DRGs. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows the induction of MANF in injured DRGs (data presented as mean ± SE; standard ‘t’ test; n=3). (C) Co-immunostaining of IB4 (yellow arrows), NF200 (white arrows) and MANF (red arrows) in normal and injured DRGs shows the preferential expression of MANF in IB4 positive non-peptidergic sensory neurons (yellow arrows in the merged view) (scale bar, 50µm).
    Figure Legend Snippet: (A) Western blot shows the expression of MANF in the normal and injured (3-day transection injury) adult rat DRGs. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows the induction of MANF in injured DRGs (data presented as mean ± SE; standard ‘t’ test; n=3). (C) Co-immunostaining of IB4 (yellow arrows), NF200 (white arrows) and MANF (red arrows) in normal and injured DRGs shows the preferential expression of MANF in IB4 positive non-peptidergic sensory neurons (yellow arrows in the merged view) (scale bar, 50µm).

    Techniques Used: Western Blot, Expressing, Control, Immunostaining

    (A) Western blot shows the expression of MANF in normal and injured (3-day transection injury) proximal and distal sciatic nerve segments of adult rats. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows no significant change in the expression of MANF in injured nerves compared to the control (data presented as mean ± SE; One-Way ANOVA; n=3). (C) Co- immunostaining of βIII tubulin and MANF in normal and injured distal sciatic nerve of adult rats shows MANF expression in a subpopulation of βIII tubulin stained axons (yellow arrows in the merged view) (scale bar, 100µm). (D) Co-immunostaining of NF200 and MANF in normal and injured distal sciatic nerve of adult rats shows no remarkable co-localization of MANF with NF200 positive axons (scale bar, 100µm). (E) Co-immunostaining of GFAP and MANF in normal and injured distal sciatic nerve of adult rats shows expression of MANF in a few populations of SCs (yellow arrows in the merged view) (scale bar, 100µm).
    Figure Legend Snippet: (A) Western blot shows the expression of MANF in normal and injured (3-day transection injury) proximal and distal sciatic nerve segments of adult rats. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows no significant change in the expression of MANF in injured nerves compared to the control (data presented as mean ± SE; One-Way ANOVA; n=3). (C) Co- immunostaining of βIII tubulin and MANF in normal and injured distal sciatic nerve of adult rats shows MANF expression in a subpopulation of βIII tubulin stained axons (yellow arrows in the merged view) (scale bar, 100µm). (D) Co-immunostaining of NF200 and MANF in normal and injured distal sciatic nerve of adult rats shows no remarkable co-localization of MANF with NF200 positive axons (scale bar, 100µm). (E) Co-immunostaining of GFAP and MANF in normal and injured distal sciatic nerve of adult rats shows expression of MANF in a few populations of SCs (yellow arrows in the merged view) (scale bar, 100µm).

    Techniques Used: Western Blot, Expressing, Control, Immunostaining, Staining

    (A) βIII tubulin staining shows that the supplementation of 50 and 100 ng/ml MANF promotes neurite outgrowth of injured and normal adult primary sensory neurons, respectively (scale bar, 100µm). (B) Quantification of neurite outgrowth using WIS-NeuroMath Software shows significant induction of neurite outgrowth in both normal and injured βIII tubulin positive neurons after MANF supplementation (data presented as mean ± SE; Standard ‘t’ test; n=3; **p<0.01, ***p<0.001). (C) NF200 staining shows that the supplementation of 50 and 100 ng/ml MANF promotes neurite outgrowth of injured and normal adult primary sensory neurons, respectively (scale bar, 100µm). (D) Quantification of neurite outgrowth using WIS-NeuroMath Software shows significant induction of neurite outgrowth in both normal and injured NF200 positive neurons after MANF supplementation (data presented as mean ± SE; Standard ‘t’ test; n=4; *p<0.05, **p<0.01).
    Figure Legend Snippet: (A) βIII tubulin staining shows that the supplementation of 50 and 100 ng/ml MANF promotes neurite outgrowth of injured and normal adult primary sensory neurons, respectively (scale bar, 100µm). (B) Quantification of neurite outgrowth using WIS-NeuroMath Software shows significant induction of neurite outgrowth in both normal and injured βIII tubulin positive neurons after MANF supplementation (data presented as mean ± SE; Standard ‘t’ test; n=3; **p<0.01, ***p<0.001). (C) NF200 staining shows that the supplementation of 50 and 100 ng/ml MANF promotes neurite outgrowth of injured and normal adult primary sensory neurons, respectively (scale bar, 100µm). (D) Quantification of neurite outgrowth using WIS-NeuroMath Software shows significant induction of neurite outgrowth in both normal and injured NF200 positive neurons after MANF supplementation (data presented as mean ± SE; Standard ‘t’ test; n=4; *p<0.05, **p<0.01).

    Techniques Used: Staining, Software

    (A) Immunostaining of GFAP in adult rat primary SCs (scale bar, 100µm). (B) Immunostaining of MANF in adult rat primary SCs. Yellow arrows show intense expression, and white arrows show weak to null expression (scale bar, 100µm). (C) MTT assay using adult rat primary SCs shows increased proliferation of cells in response to exogenous MANF supplementation (data presented as mean ± SE; One-Way ANOVA (Tukey’s multiple comparisons test); n=6; *p<0.05). (D) Brightfield images of the scratch assay using adult rat primary SCs show a time-dependent gap closure in control and MANF (50 ng/ml) supplemented groups (scale bar, 100µm). (E) Quantification of percentage gap closure in scratch assay shows increased gap closure in MANF (50 ng/ml) supplemented group compared to control (data presented as mean ± SE; Standard ‘t’ test; n=3; *p<0.05, **p<0.01). (F) Schematic of transwell migration assay. (G) DAPI staining on the lower side of the membrane from a transwell migration assay shows migrated adult primary SCs (scale bar, 100µm). (H) Quantification of transwell migration assay shows increased migration of primary SCs in MANF (50 ng/ml) supplemented group (data presented as mean ± SE; standard ‘t’ test; n=3; **p<0.01).
    Figure Legend Snippet: (A) Immunostaining of GFAP in adult rat primary SCs (scale bar, 100µm). (B) Immunostaining of MANF in adult rat primary SCs. Yellow arrows show intense expression, and white arrows show weak to null expression (scale bar, 100µm). (C) MTT assay using adult rat primary SCs shows increased proliferation of cells in response to exogenous MANF supplementation (data presented as mean ± SE; One-Way ANOVA (Tukey’s multiple comparisons test); n=6; *p<0.05). (D) Brightfield images of the scratch assay using adult rat primary SCs show a time-dependent gap closure in control and MANF (50 ng/ml) supplemented groups (scale bar, 100µm). (E) Quantification of percentage gap closure in scratch assay shows increased gap closure in MANF (50 ng/ml) supplemented group compared to control (data presented as mean ± SE; Standard ‘t’ test; n=3; *p<0.05, **p<0.01). (F) Schematic of transwell migration assay. (G) DAPI staining on the lower side of the membrane from a transwell migration assay shows migrated adult primary SCs (scale bar, 100µm). (H) Quantification of transwell migration assay shows increased migration of primary SCs in MANF (50 ng/ml) supplemented group (data presented as mean ± SE; standard ‘t’ test; n=3; **p<0.01).

    Techniques Used: Immunostaining, Expressing, MTT Assay, Wound Healing Assay, Control, Transwell Migration Assay, Staining, Membrane, Migration

    (A) MTT assay using S16 SC line shows increased proliferation of cells in response to exogenous MANF (data presented as mean ± SE; One-Way ANOVA (Tukey’s multiple comparisons test); n=8; **p<0.01, ****p<0.0001). (B) DAPI staining on the lower side of the membrane from a transwell migration assay shows migrated S16 SCs at 48h in the control and MANF group (scale bar, 200µm). (C) Quantification of transwell migration assay shows increased migration of S16 SCs in MANF (100 ng/ml) supplemented group at 48h (data presented as mean ± SE; standard ‘t’ test; n=3; *p<0.05).
    Figure Legend Snippet: (A) MTT assay using S16 SC line shows increased proliferation of cells in response to exogenous MANF (data presented as mean ± SE; One-Way ANOVA (Tukey’s multiple comparisons test); n=8; **p<0.01, ****p<0.0001). (B) DAPI staining on the lower side of the membrane from a transwell migration assay shows migrated S16 SCs at 48h in the control and MANF group (scale bar, 200µm). (C) Quantification of transwell migration assay shows increased migration of S16 SCs in MANF (100 ng/ml) supplemented group at 48h (data presented as mean ± SE; standard ‘t’ test; n=3; *p<0.05).

    Techniques Used: MTT Assay, Staining, Membrane, Transwell Migration Assay, Control, Migration

    (A) Schematic of in vivo nerve crush injury and local supplementation of MANF in adult mice. (B) Schematic of axon quantification approach. The number of axons crossing the perpendicular lines drawn at the shown distances are manually counted to tabulate the axon count at each distance. (C) βIII tubulin staining of sections of injured sciatic nerves from adult mice shows degenerated (blue arrows) and intact (green arrows) axons in the control and MANF group, respectively. White arrows show the crush site (scale bar, 200µm). (D) Quantification of intact axons in the distal nerve segment shows an increased number of axons in the MANF group compared to the control at varying distances from the crush site (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; **p<0.01).
    Figure Legend Snippet: (A) Schematic of in vivo nerve crush injury and local supplementation of MANF in adult mice. (B) Schematic of axon quantification approach. The number of axons crossing the perpendicular lines drawn at the shown distances are manually counted to tabulate the axon count at each distance. (C) βIII tubulin staining of sections of injured sciatic nerves from adult mice shows degenerated (blue arrows) and intact (green arrows) axons in the control and MANF group, respectively. White arrows show the crush site (scale bar, 200µm). (D) Quantification of intact axons in the distal nerve segment shows an increased number of axons in the MANF group compared to the control at varying distances from the crush site (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; **p<0.01).

    Techniques Used: In Vivo, Staining, Control

    Maps of (A) pLVX TetOne Puro (empty vector) and (B) pLVX TetOne Puro MANF constructs.
    Figure Legend Snippet: Maps of (A) pLVX TetOne Puro (empty vector) and (B) pLVX TetOne Puro MANF constructs.

    Techniques Used: Plasmid Preparation, Construct

    (A) SC-MANF after seven days of puromycin selection in a culture flask. (B ) SC- MANF grows healthy on an artificial nerve conduit (15-day). (C) ELISA assay shows increased secretion of MANF in the SC-MANF group compared to L-EV transduced primary SCs after Dox supplementation (data presented as mean ± SE; standard ‘t’ test; n=4). (D) Schematic of nerve crush injury experiment using DRG-nerve explant. (E) Immunostaining of βIII tubulin in sections of crush injured DRG-nerve explant (day 6) showing intact axons (yellow arrows) in the L-EV and L-MANF transduced groups. The white arrow shows DRG (scale bar, 200µm). (F) Quantification of axons in DRG-nerve explants on day 6 shows an increased number of intact axons in the L- MANF group compared to the L-EV group (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; *p<0.05, **p<0.01, ***p<0.001). (G) Immunostaining shows the expression of MANF in L-EV and L-MANF transduced DRG-nerve explants cultured in the presence of Dox (1µg/ml). White arrows show the expression of MANF (green dots) in the nerve segment, and the red arrow shows MANF expression in DRG (scale bar, 500µm). Enlarged areas are provided in the insets. (H) A representative image shows the expression of MANF in SCs (SC-MANF generation) in the nerve segment of a DRG-nerve explant transduced with L-MANF and cultured in the presence of Dox.
    Figure Legend Snippet: (A) SC-MANF after seven days of puromycin selection in a culture flask. (B ) SC- MANF grows healthy on an artificial nerve conduit (15-day). (C) ELISA assay shows increased secretion of MANF in the SC-MANF group compared to L-EV transduced primary SCs after Dox supplementation (data presented as mean ± SE; standard ‘t’ test; n=4). (D) Schematic of nerve crush injury experiment using DRG-nerve explant. (E) Immunostaining of βIII tubulin in sections of crush injured DRG-nerve explant (day 6) showing intact axons (yellow arrows) in the L-EV and L-MANF transduced groups. The white arrow shows DRG (scale bar, 200µm). (F) Quantification of axons in DRG-nerve explants on day 6 shows an increased number of intact axons in the L- MANF group compared to the L-EV group (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; *p<0.05, **p<0.01, ***p<0.001). (G) Immunostaining shows the expression of MANF in L-EV and L-MANF transduced DRG-nerve explants cultured in the presence of Dox (1µg/ml). White arrows show the expression of MANF (green dots) in the nerve segment, and the red arrow shows MANF expression in DRG (scale bar, 500µm). Enlarged areas are provided in the insets. (H) A representative image shows the expression of MANF in SCs (SC-MANF generation) in the nerve segment of a DRG-nerve explant transduced with L-MANF and cultured in the presence of Dox.

    Techniques Used: Selection, Enzyme-linked Immunosorbent Assay, Immunostaining, Expressing, Cell Culture, Transduction

    (A) Schematic and timeline of the nerve crush injury experiment using DRG-nerve explants. (E) Immunostaining of GAP-43 in sections of DRG-nerve explants (day 15) shows longer regenerating axons (yellow arrows) past crush site (white arrow) in the L-MANF group compared the L-EV group (scale bar, 500µm). (B) Quantification of regenerating proximal and distal axons in DRG-nerve explants on day 15 shows a significant increase in regenerating distal axons in the L-MANF group compared to L-EV group (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; *p<0.05, **p<0.01). (C) GFAP staining in sections of DRG-nerve explants (day 15) shows healthy distribution of SCs in the distal nerve of L-MANF group (scale bar, 500µm). (D) Immunostaining shows the expression of MANF in L-EV and L-MANF transduced DRG-nerve explants cultured in the presence of Dox (1µg/ml) for 15 days. Yellow arrows show the expression of MANF (green dots) in the nerve segment (scale bar, 100µm).
    Figure Legend Snippet: (A) Schematic and timeline of the nerve crush injury experiment using DRG-nerve explants. (E) Immunostaining of GAP-43 in sections of DRG-nerve explants (day 15) shows longer regenerating axons (yellow arrows) past crush site (white arrow) in the L-MANF group compared the L-EV group (scale bar, 500µm). (B) Quantification of regenerating proximal and distal axons in DRG-nerve explants on day 15 shows a significant increase in regenerating distal axons in the L-MANF group compared to L-EV group (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; *p<0.05, **p<0.01). (C) GFAP staining in sections of DRG-nerve explants (day 15) shows healthy distribution of SCs in the distal nerve of L-MANF group (scale bar, 500µm). (D) Immunostaining shows the expression of MANF in L-EV and L-MANF transduced DRG-nerve explants cultured in the presence of Dox (1µg/ml) for 15 days. Yellow arrows show the expression of MANF (green dots) in the nerve segment (scale bar, 100µm).

    Techniques Used: Immunostaining, Staining, Expressing, Cell Culture



    Similar Products

    manf elisa kit  (Biomatik)
    90
    Biomatik manf elisa kit
    (A) Western blot shows the expression of <t>MANF</t> in the normal and injured (3-day transection injury) adult rat DRGs. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows the induction of MANF in injured DRGs (data presented as mean ± SE; standard ‘t’ test; n=3). (C) Co-immunostaining of IB4 (yellow arrows), NF200 (white arrows) and MANF (red arrows) in normal and injured DRGs shows the preferential expression of MANF in IB4 positive non-peptidergic sensory neurons (yellow arrows in the merged view) (scale bar, 50µm).
    Manf Elisa Kit, supplied by Biomatik, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/bio_rxiv__2025__03__14__642904-110-7-14?v=Biomatik
    Average 90 stars, based on 1 article reviews
    manf elisa kit - by Bioz Stars, 2026-07
    90/100 stars
    		  Buy from Supplier
    human manf elisa kit  (Cloud-Clone corp)
    90
    Cloud-Clone corp human manf elisa kit
    (A) Western blot shows the expression of <t>MANF</t> in the normal and injured (3-day transection injury) adult rat DRGs. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows the induction of MANF in injured DRGs (data presented as mean ± SE; standard ‘t’ test; n=3). (C) Co-immunostaining of IB4 (yellow arrows), NF200 (white arrows) and MANF (red arrows) in normal and injured DRGs shows the preferential expression of MANF in IB4 positive non-peptidergic sensory neurons (yellow arrows in the merged view) (scale bar, 50µm).
    Human Manf Elisa Kit, supplied by Cloud-Clone corp, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/pm38493745-99-15-19?v=Cloud-Clone+corp
    Average 90 stars, based on 1 article reviews
    human manf elisa kit - by Bioz Stars, 2026-07
    90/100 stars
    		  Buy from Supplier
    human manf picokine elisa kits  (Boster Bio)
    91
    Boster Bio human manf picokine elisa kits
    Fig. 2 Sex-specific scatter plot graphs showing the association of IL-33 and <t>MANF</t> levels with Ham-D scores of study participants
    Human Manf Picokine Elisa Kits, supplied by Boster Bio, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/pm38216957-87-8-13?v=Boster+Bio
    Average 91 stars, based on 1 article reviews
    human manf picokine elisa kits - by Bioz Stars, 2026-07
    91/100 stars
    		  Buy from Supplier
    manf elisa kit  (Reddot Biotech)
    90
    Reddot Biotech manf elisa kit
    Fig. 2 Sex-specific scatter plot graphs showing the association of IL-33 and <t>MANF</t> levels with Ham-D scores of study participants
    Manf Elisa Kit, supplied by Reddot Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/pm37652307-51-13-16?v=Reddot+Biotech
    Average 90 stars, based on 1 article reviews
    manf elisa kit - by Bioz Stars, 2026-07
    90/100 stars
    		  Buy from Supplier
    mouse manf elisa kit  (Cloud-Clone corp)
    90
    Cloud-Clone corp mouse manf elisa kit
    <t>MANF</t> expression is upregulated in fibrotic liver tissues. (A) MANF level in hepatic tissues of the patients with hepatitis ( n = 8), hepatic fibrosis ( n = 7), and hepatic cirrhosis ( n = 7) detected by using immunohistochemistry assay, and the integral optical density were calculated. (B) HE, Sirius red, and Masson staining in CCl 4 -treated fibrotic liver tissues were performed, and the positive area was calculated. MANF was detected by immunohistochemistry and the integral optical density was calculated ( n = 10). (C, D) MANF protein (C) and mRNA (D) levels in CCl 4 -treated fibrotic liver tissues were detected by Western blot and qPCR ( n = 10), respectively. The relative MANF levels were calculated. (E) Serum MANF was detected by <t>ELISA.</t> (F) Double labeled immunofluorescence was performed to detect the colocalization of MANF (red) with α -SMA (green), HNF-4 (green), F4/80 (green), and Ly6G (green) in fibrotic liver tissues ( n = 10). DAPI (blue) was used to stain nuclei. Data are expressed as mean ± SEM. ∗∗∗ P < 0.001. WT, wild type; CCl 4 , carbon tetrachloride.
    Mouse Manf Elisa Kit, supplied by Cloud-Clone corp, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/pmc10547964-69-59-63?v=Cloud-Clone+corp
    Average 90 stars, based on 1 article reviews
    mouse manf elisa kit - by Bioz Stars, 2026-07
    90/100 stars
    		  Buy from Supplier
    manf elisa kit  (Cloud-Clone corp)
    90
    Cloud-Clone corp manf elisa kit
    <t>MANF</t> expression is upregulated in fibrotic liver tissues. (A) MANF level in hepatic tissues of the patients with hepatitis ( n = 8), hepatic fibrosis ( n = 7), and hepatic cirrhosis ( n = 7) detected by using immunohistochemistry assay, and the integral optical density were calculated. (B) HE, Sirius red, and Masson staining in CCl 4 -treated fibrotic liver tissues were performed, and the positive area was calculated. MANF was detected by immunohistochemistry and the integral optical density was calculated ( n = 10). (C, D) MANF protein (C) and mRNA (D) levels in CCl 4 -treated fibrotic liver tissues were detected by Western blot and qPCR ( n = 10), respectively. The relative MANF levels were calculated. (E) Serum MANF was detected by <t>ELISA.</t> (F) Double labeled immunofluorescence was performed to detect the colocalization of MANF (red) with α -SMA (green), HNF-4 (green), F4/80 (green), and Ly6G (green) in fibrotic liver tissues ( n = 10). DAPI (blue) was used to stain nuclei. Data are expressed as mean ± SEM. ∗∗∗ P < 0.001. WT, wild type; CCl 4 , carbon tetrachloride.
    Manf Elisa Kit, supplied by Cloud-Clone corp, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/pm36689049-61-14-17?v=Cloud-Clone+corp
    Average 90 stars, based on 1 article reviews
    manf elisa kit - by Bioz Stars, 2026-07
    90/100 stars
    		  Buy from Supplier
    human manf elisa kit  (Boster Bio)
    90
    Boster Bio human manf elisa kit
    <t>MANF</t> expression is upregulated in fibrotic liver tissues. (A) MANF level in hepatic tissues of the patients with hepatitis ( n = 8), hepatic fibrosis ( n = 7), and hepatic cirrhosis ( n = 7) detected by using immunohistochemistry assay, and the integral optical density were calculated. (B) HE, Sirius red, and Masson staining in CCl 4 -treated fibrotic liver tissues were performed, and the positive area was calculated. MANF was detected by immunohistochemistry and the integral optical density was calculated ( n = 10). (C, D) MANF protein (C) and mRNA (D) levels in CCl 4 -treated fibrotic liver tissues were detected by Western blot and qPCR ( n = 10), respectively. The relative MANF levels were calculated. (E) Serum MANF was detected by <t>ELISA.</t> (F) Double labeled immunofluorescence was performed to detect the colocalization of MANF (red) with α -SMA (green), HNF-4 (green), F4/80 (green), and Ly6G (green) in fibrotic liver tissues ( n = 10). DAPI (blue) was used to stain nuclei. Data are expressed as mean ± SEM. ∗∗∗ P < 0.001. WT, wild type; CCl 4 , carbon tetrachloride.
    Human Manf Elisa Kit, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/pm36087374-69-17-21?v=Boster+Bio
    Average 90 stars, based on 1 article reviews
    human manf elisa kit - by Bioz Stars, 2026-07
    90/100 stars
    		  Buy from Supplier
    rat manf elisa kit  (Boster Bio)
    93
    Boster Bio rat manf elisa kit
    <t>MANF</t> expression is upregulated in fibrotic liver tissues. (A) MANF level in hepatic tissues of the patients with hepatitis ( n = 8), hepatic fibrosis ( n = 7), and hepatic cirrhosis ( n = 7) detected by using immunohistochemistry assay, and the integral optical density were calculated. (B) HE, Sirius red, and Masson staining in CCl 4 -treated fibrotic liver tissues were performed, and the positive area was calculated. MANF was detected by immunohistochemistry and the integral optical density was calculated ( n = 10). (C, D) MANF protein (C) and mRNA (D) levels in CCl 4 -treated fibrotic liver tissues were detected by Western blot and qPCR ( n = 10), respectively. The relative MANF levels were calculated. (E) Serum MANF was detected by <t>ELISA.</t> (F) Double labeled immunofluorescence was performed to detect the colocalization of MANF (red) with α -SMA (green), HNF-4 (green), F4/80 (green), and Ly6G (green) in fibrotic liver tissues ( n = 10). DAPI (blue) was used to stain nuclei. Data are expressed as mean ± SEM. ∗∗∗ P < 0.001. WT, wild type; CCl 4 , carbon tetrachloride.
    Rat Manf Elisa Kit, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/pm36087374-69-26-21?v=Boster+Bio
    Average 93 stars, based on 1 article reviews
    rat manf elisa kit - by Bioz Stars, 2026-07
    93/100 stars
    		  Buy from Supplier
    manf elisa kit  (Sino Biological)
    90
    Sino Biological manf elisa kit
    <t>MANF</t> expression is upregulated in fibrotic liver tissues. (A) MANF level in hepatic tissues of the patients with hepatitis ( n = 8), hepatic fibrosis ( n = 7), and hepatic cirrhosis ( n = 7) detected by using immunohistochemistry assay, and the integral optical density were calculated. (B) HE, Sirius red, and Masson staining in CCl 4 -treated fibrotic liver tissues were performed, and the positive area was calculated. MANF was detected by immunohistochemistry and the integral optical density was calculated ( n = 10). (C, D) MANF protein (C) and mRNA (D) levels in CCl 4 -treated fibrotic liver tissues were detected by Western blot and qPCR ( n = 10), respectively. The relative MANF levels were calculated. (E) Serum MANF was detected by <t>ELISA.</t> (F) Double labeled immunofluorescence was performed to detect the colocalization of MANF (red) with α -SMA (green), HNF-4 (green), F4/80 (green), and Ly6G (green) in fibrotic liver tissues ( n = 10). DAPI (blue) was used to stain nuclei. Data are expressed as mean ± SEM. ∗∗∗ P < 0.001. WT, wild type; CCl 4 , carbon tetrachloride.
    Manf Elisa Kit, supplied by Sino Biological, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/manf+elisa+kit/pmc09515611-54-12-16?v=Sino+Biological
    Average 90 stars, based on 1 article reviews
    manf elisa kit - by Bioz Stars, 2026-07
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) Western blot shows the expression of MANF in the normal and injured (3-day transection injury) adult rat DRGs. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows the induction of MANF in injured DRGs (data presented as mean ± SE; standard ‘t’ test; n=3). (C) Co-immunostaining of IB4 (yellow arrows), NF200 (white arrows) and MANF (red arrows) in normal and injured DRGs shows the preferential expression of MANF in IB4 positive non-peptidergic sensory neurons (yellow arrows in the merged view) (scale bar, 50µm).

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: (A) Western blot shows the expression of MANF in the normal and injured (3-day transection injury) adult rat DRGs. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows the induction of MANF in injured DRGs (data presented as mean ± SE; standard ‘t’ test; n=3). (C) Co-immunostaining of IB4 (yellow arrows), NF200 (white arrows) and MANF (red arrows) in normal and injured DRGs shows the preferential expression of MANF in IB4 positive non-peptidergic sensory neurons (yellow arrows in the merged view) (scale bar, 50µm).

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: Western Blot, Expressing, Control, Immunostaining

    (A) Western blot shows the expression of MANF in normal and injured (3-day transection injury) proximal and distal sciatic nerve segments of adult rats. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows no significant change in the expression of MANF in injured nerves compared to the control (data presented as mean ± SE; One-Way ANOVA; n=3). (C) Co- immunostaining of βIII tubulin and MANF in normal and injured distal sciatic nerve of adult rats shows MANF expression in a subpopulation of βIII tubulin stained axons (yellow arrows in the merged view) (scale bar, 100µm). (D) Co-immunostaining of NF200 and MANF in normal and injured distal sciatic nerve of adult rats shows no remarkable co-localization of MANF with NF200 positive axons (scale bar, 100µm). (E) Co-immunostaining of GFAP and MANF in normal and injured distal sciatic nerve of adult rats shows expression of MANF in a few populations of SCs (yellow arrows in the merged view) (scale bar, 100µm).

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: (A) Western blot shows the expression of MANF in normal and injured (3-day transection injury) proximal and distal sciatic nerve segments of adult rats. GAPDH is used as the loading control. (B) Quantification of ‘A’ shows no significant change in the expression of MANF in injured nerves compared to the control (data presented as mean ± SE; One-Way ANOVA; n=3). (C) Co- immunostaining of βIII tubulin and MANF in normal and injured distal sciatic nerve of adult rats shows MANF expression in a subpopulation of βIII tubulin stained axons (yellow arrows in the merged view) (scale bar, 100µm). (D) Co-immunostaining of NF200 and MANF in normal and injured distal sciatic nerve of adult rats shows no remarkable co-localization of MANF with NF200 positive axons (scale bar, 100µm). (E) Co-immunostaining of GFAP and MANF in normal and injured distal sciatic nerve of adult rats shows expression of MANF in a few populations of SCs (yellow arrows in the merged view) (scale bar, 100µm).

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: Western Blot, Expressing, Control, Immunostaining, Staining

    (A) βIII tubulin staining shows that the supplementation of 50 and 100 ng/ml MANF promotes neurite outgrowth of injured and normal adult primary sensory neurons, respectively (scale bar, 100µm). (B) Quantification of neurite outgrowth using WIS-NeuroMath Software shows significant induction of neurite outgrowth in both normal and injured βIII tubulin positive neurons after MANF supplementation (data presented as mean ± SE; Standard ‘t’ test; n=3; **p<0.01, ***p<0.001). (C) NF200 staining shows that the supplementation of 50 and 100 ng/ml MANF promotes neurite outgrowth of injured and normal adult primary sensory neurons, respectively (scale bar, 100µm). (D) Quantification of neurite outgrowth using WIS-NeuroMath Software shows significant induction of neurite outgrowth in both normal and injured NF200 positive neurons after MANF supplementation (data presented as mean ± SE; Standard ‘t’ test; n=4; *p<0.05, **p<0.01).

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: (A) βIII tubulin staining shows that the supplementation of 50 and 100 ng/ml MANF promotes neurite outgrowth of injured and normal adult primary sensory neurons, respectively (scale bar, 100µm). (B) Quantification of neurite outgrowth using WIS-NeuroMath Software shows significant induction of neurite outgrowth in both normal and injured βIII tubulin positive neurons after MANF supplementation (data presented as mean ± SE; Standard ‘t’ test; n=3; **p<0.01, ***p<0.001). (C) NF200 staining shows that the supplementation of 50 and 100 ng/ml MANF promotes neurite outgrowth of injured and normal adult primary sensory neurons, respectively (scale bar, 100µm). (D) Quantification of neurite outgrowth using WIS-NeuroMath Software shows significant induction of neurite outgrowth in both normal and injured NF200 positive neurons after MANF supplementation (data presented as mean ± SE; Standard ‘t’ test; n=4; *p<0.05, **p<0.01).

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: Staining, Software

    (A) Immunostaining of GFAP in adult rat primary SCs (scale bar, 100µm). (B) Immunostaining of MANF in adult rat primary SCs. Yellow arrows show intense expression, and white arrows show weak to null expression (scale bar, 100µm). (C) MTT assay using adult rat primary SCs shows increased proliferation of cells in response to exogenous MANF supplementation (data presented as mean ± SE; One-Way ANOVA (Tukey’s multiple comparisons test); n=6; *p<0.05). (D) Brightfield images of the scratch assay using adult rat primary SCs show a time-dependent gap closure in control and MANF (50 ng/ml) supplemented groups (scale bar, 100µm). (E) Quantification of percentage gap closure in scratch assay shows increased gap closure in MANF (50 ng/ml) supplemented group compared to control (data presented as mean ± SE; Standard ‘t’ test; n=3; *p<0.05, **p<0.01). (F) Schematic of transwell migration assay. (G) DAPI staining on the lower side of the membrane from a transwell migration assay shows migrated adult primary SCs (scale bar, 100µm). (H) Quantification of transwell migration assay shows increased migration of primary SCs in MANF (50 ng/ml) supplemented group (data presented as mean ± SE; standard ‘t’ test; n=3; **p<0.01).

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: (A) Immunostaining of GFAP in adult rat primary SCs (scale bar, 100µm). (B) Immunostaining of MANF in adult rat primary SCs. Yellow arrows show intense expression, and white arrows show weak to null expression (scale bar, 100µm). (C) MTT assay using adult rat primary SCs shows increased proliferation of cells in response to exogenous MANF supplementation (data presented as mean ± SE; One-Way ANOVA (Tukey’s multiple comparisons test); n=6; *p<0.05). (D) Brightfield images of the scratch assay using adult rat primary SCs show a time-dependent gap closure in control and MANF (50 ng/ml) supplemented groups (scale bar, 100µm). (E) Quantification of percentage gap closure in scratch assay shows increased gap closure in MANF (50 ng/ml) supplemented group compared to control (data presented as mean ± SE; Standard ‘t’ test; n=3; *p<0.05, **p<0.01). (F) Schematic of transwell migration assay. (G) DAPI staining on the lower side of the membrane from a transwell migration assay shows migrated adult primary SCs (scale bar, 100µm). (H) Quantification of transwell migration assay shows increased migration of primary SCs in MANF (50 ng/ml) supplemented group (data presented as mean ± SE; standard ‘t’ test; n=3; **p<0.01).

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: Immunostaining, Expressing, MTT Assay, Wound Healing Assay, Control, Transwell Migration Assay, Staining, Membrane, Migration

    (A) MTT assay using S16 SC line shows increased proliferation of cells in response to exogenous MANF (data presented as mean ± SE; One-Way ANOVA (Tukey’s multiple comparisons test); n=8; **p<0.01, ****p<0.0001). (B) DAPI staining on the lower side of the membrane from a transwell migration assay shows migrated S16 SCs at 48h in the control and MANF group (scale bar, 200µm). (C) Quantification of transwell migration assay shows increased migration of S16 SCs in MANF (100 ng/ml) supplemented group at 48h (data presented as mean ± SE; standard ‘t’ test; n=3; *p<0.05).

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: (A) MTT assay using S16 SC line shows increased proliferation of cells in response to exogenous MANF (data presented as mean ± SE; One-Way ANOVA (Tukey’s multiple comparisons test); n=8; **p<0.01, ****p<0.0001). (B) DAPI staining on the lower side of the membrane from a transwell migration assay shows migrated S16 SCs at 48h in the control and MANF group (scale bar, 200µm). (C) Quantification of transwell migration assay shows increased migration of S16 SCs in MANF (100 ng/ml) supplemented group at 48h (data presented as mean ± SE; standard ‘t’ test; n=3; *p<0.05).

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: MTT Assay, Staining, Membrane, Transwell Migration Assay, Control, Migration

    (A) Schematic of in vivo nerve crush injury and local supplementation of MANF in adult mice. (B) Schematic of axon quantification approach. The number of axons crossing the perpendicular lines drawn at the shown distances are manually counted to tabulate the axon count at each distance. (C) βIII tubulin staining of sections of injured sciatic nerves from adult mice shows degenerated (blue arrows) and intact (green arrows) axons in the control and MANF group, respectively. White arrows show the crush site (scale bar, 200µm). (D) Quantification of intact axons in the distal nerve segment shows an increased number of axons in the MANF group compared to the control at varying distances from the crush site (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; **p<0.01).

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: (A) Schematic of in vivo nerve crush injury and local supplementation of MANF in adult mice. (B) Schematic of axon quantification approach. The number of axons crossing the perpendicular lines drawn at the shown distances are manually counted to tabulate the axon count at each distance. (C) βIII tubulin staining of sections of injured sciatic nerves from adult mice shows degenerated (blue arrows) and intact (green arrows) axons in the control and MANF group, respectively. White arrows show the crush site (scale bar, 200µm). (D) Quantification of intact axons in the distal nerve segment shows an increased number of axons in the MANF group compared to the control at varying distances from the crush site (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; **p<0.01).

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: In Vivo, Staining, Control

    Maps of (A) pLVX TetOne Puro (empty vector) and (B) pLVX TetOne Puro MANF constructs.

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: Maps of (A) pLVX TetOne Puro (empty vector) and (B) pLVX TetOne Puro MANF constructs.

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: Plasmid Preparation, Construct

    (A) SC-MANF after seven days of puromycin selection in a culture flask. (B ) SC- MANF grows healthy on an artificial nerve conduit (15-day). (C) ELISA assay shows increased secretion of MANF in the SC-MANF group compared to L-EV transduced primary SCs after Dox supplementation (data presented as mean ± SE; standard ‘t’ test; n=4). (D) Schematic of nerve crush injury experiment using DRG-nerve explant. (E) Immunostaining of βIII tubulin in sections of crush injured DRG-nerve explant (day 6) showing intact axons (yellow arrows) in the L-EV and L-MANF transduced groups. The white arrow shows DRG (scale bar, 200µm). (F) Quantification of axons in DRG-nerve explants on day 6 shows an increased number of intact axons in the L- MANF group compared to the L-EV group (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; *p<0.05, **p<0.01, ***p<0.001). (G) Immunostaining shows the expression of MANF in L-EV and L-MANF transduced DRG-nerve explants cultured in the presence of Dox (1µg/ml). White arrows show the expression of MANF (green dots) in the nerve segment, and the red arrow shows MANF expression in DRG (scale bar, 500µm). Enlarged areas are provided in the insets. (H) A representative image shows the expression of MANF in SCs (SC-MANF generation) in the nerve segment of a DRG-nerve explant transduced with L-MANF and cultured in the presence of Dox.

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: (A) SC-MANF after seven days of puromycin selection in a culture flask. (B ) SC- MANF grows healthy on an artificial nerve conduit (15-day). (C) ELISA assay shows increased secretion of MANF in the SC-MANF group compared to L-EV transduced primary SCs after Dox supplementation (data presented as mean ± SE; standard ‘t’ test; n=4). (D) Schematic of nerve crush injury experiment using DRG-nerve explant. (E) Immunostaining of βIII tubulin in sections of crush injured DRG-nerve explant (day 6) showing intact axons (yellow arrows) in the L-EV and L-MANF transduced groups. The white arrow shows DRG (scale bar, 200µm). (F) Quantification of axons in DRG-nerve explants on day 6 shows an increased number of intact axons in the L- MANF group compared to the L-EV group (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; *p<0.05, **p<0.01, ***p<0.001). (G) Immunostaining shows the expression of MANF in L-EV and L-MANF transduced DRG-nerve explants cultured in the presence of Dox (1µg/ml). White arrows show the expression of MANF (green dots) in the nerve segment, and the red arrow shows MANF expression in DRG (scale bar, 500µm). Enlarged areas are provided in the insets. (H) A representative image shows the expression of MANF in SCs (SC-MANF generation) in the nerve segment of a DRG-nerve explant transduced with L-MANF and cultured in the presence of Dox.

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: Selection, Enzyme-linked Immunosorbent Assay, Immunostaining, Expressing, Cell Culture, Transduction

    (A) Schematic and timeline of the nerve crush injury experiment using DRG-nerve explants. (E) Immunostaining of GAP-43 in sections of DRG-nerve explants (day 15) shows longer regenerating axons (yellow arrows) past crush site (white arrow) in the L-MANF group compared the L-EV group (scale bar, 500µm). (B) Quantification of regenerating proximal and distal axons in DRG-nerve explants on day 15 shows a significant increase in regenerating distal axons in the L-MANF group compared to L-EV group (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; *p<0.05, **p<0.01). (C) GFAP staining in sections of DRG-nerve explants (day 15) shows healthy distribution of SCs in the distal nerve of L-MANF group (scale bar, 500µm). (D) Immunostaining shows the expression of MANF in L-EV and L-MANF transduced DRG-nerve explants cultured in the presence of Dox (1µg/ml) for 15 days. Yellow arrows show the expression of MANF (green dots) in the nerve segment (scale bar, 100µm).

    Journal: bioRxiv

    Article Title: Schwann cells modified to secrete MANF is a potential cellular therapy for peripheral nerve regeneration

    doi: 10.1101/2025.03.14.642904

    Figure Lengend Snippet: (A) Schematic and timeline of the nerve crush injury experiment using DRG-nerve explants. (E) Immunostaining of GAP-43 in sections of DRG-nerve explants (day 15) shows longer regenerating axons (yellow arrows) past crush site (white arrow) in the L-MANF group compared the L-EV group (scale bar, 500µm). (B) Quantification of regenerating proximal and distal axons in DRG-nerve explants on day 15 shows a significant increase in regenerating distal axons in the L-MANF group compared to L-EV group (data presented as mean ± SE; Two-Way ANOVA (Sidak’s multiple comparisons test); n=3; *p<0.05, **p<0.01). (C) GFAP staining in sections of DRG-nerve explants (day 15) shows healthy distribution of SCs in the distal nerve of L-MANF group (scale bar, 500µm). (D) Immunostaining shows the expression of MANF in L-EV and L-MANF transduced DRG-nerve explants cultured in the presence of Dox (1µg/ml) for 15 days. Yellow arrows show the expression of MANF (green dots) in the nerve segment (scale bar, 100µm).

    Article Snippet: ELISA was performed using the commercially available MANF ELISA Kit (Cat No. EKL58888- 96T, Biomatik) according to the manufacturer’s instructions.

    Techniques: Immunostaining, Staining, Expressing, Cell Culture

    Fig. 2 Sex-specific scatter plot graphs showing the association of IL-33 and MANF levels with Ham-D scores of study participants

    Journal: BMC psychiatry

    Article Title: Serum levels of interleukin-33 and mesencephalic astrocyte derived neurotrophic factors in patients with major depressive disorder: a cross-sectional comparative design.

    doi: 10.1186/s12888-023-05463-8

    Figure Lengend Snippet: Fig. 2 Sex-specific scatter plot graphs showing the association of IL-33 and MANF levels with Ham-D scores of study participants

    Article Snippet: Commercially available Human IL-33 Picokine ELISA kits and Human MANF PicoKine ELISA kits (Boster Bio, USA) were used to determine the serum levels of IL-33 and MANF, respectively, for further analysis.

    Techniques:

    MANF expression is upregulated in fibrotic liver tissues. (A) MANF level in hepatic tissues of the patients with hepatitis ( n = 8), hepatic fibrosis ( n = 7), and hepatic cirrhosis ( n = 7) detected by using immunohistochemistry assay, and the integral optical density were calculated. (B) HE, Sirius red, and Masson staining in CCl 4 -treated fibrotic liver tissues were performed, and the positive area was calculated. MANF was detected by immunohistochemistry and the integral optical density was calculated ( n = 10). (C, D) MANF protein (C) and mRNA (D) levels in CCl 4 -treated fibrotic liver tissues were detected by Western blot and qPCR ( n = 10), respectively. The relative MANF levels were calculated. (E) Serum MANF was detected by ELISA. (F) Double labeled immunofluorescence was performed to detect the colocalization of MANF (red) with α -SMA (green), HNF-4 (green), F4/80 (green), and Ly6G (green) in fibrotic liver tissues ( n = 10). DAPI (blue) was used to stain nuclei. Data are expressed as mean ± SEM. ∗∗∗ P < 0.001. WT, wild type; CCl 4 , carbon tetrachloride.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: MANF brakes TLR4 signaling by competitively binding S100A8 with S100A9 to regulate macrophage phenotypes in hepatic fibrosis

    doi: 10.1016/j.apsb.2023.07.027

    Figure Lengend Snippet: MANF expression is upregulated in fibrotic liver tissues. (A) MANF level in hepatic tissues of the patients with hepatitis ( n = 8), hepatic fibrosis ( n = 7), and hepatic cirrhosis ( n = 7) detected by using immunohistochemistry assay, and the integral optical density were calculated. (B) HE, Sirius red, and Masson staining in CCl 4 -treated fibrotic liver tissues were performed, and the positive area was calculated. MANF was detected by immunohistochemistry and the integral optical density was calculated ( n = 10). (C, D) MANF protein (C) and mRNA (D) levels in CCl 4 -treated fibrotic liver tissues were detected by Western blot and qPCR ( n = 10), respectively. The relative MANF levels were calculated. (E) Serum MANF was detected by ELISA. (F) Double labeled immunofluorescence was performed to detect the colocalization of MANF (red) with α -SMA (green), HNF-4 (green), F4/80 (green), and Ly6G (green) in fibrotic liver tissues ( n = 10). DAPI (blue) was used to stain nuclei. Data are expressed as mean ± SEM. ∗∗∗ P < 0.001. WT, wild type; CCl 4 , carbon tetrachloride.

    Article Snippet: The utilized ELISA Kits included Mouse TGF- β 1 ELISA Kit (R&D Systems, Minneapolis, USA, MB100B), Mouse TNF- α ELISA Kit (Abcam, ab208348), Mouse IL-1 β ELISA Kit (Abcam, ab197742), Mouse CCL2 ELISA Kit (Abcam, ab208979), Mouse S100A8 ELISA Kit (Abcam, ab263886), Mouse S100A9 ELISA Kit (Abcam, ab213887), Mouse Hyaluronan ELISA Kit (Jiyinmei Biological Technology, Wuhan, China, JYM0514Mo), and Mouse MANF ELISA Kit (Cloud-Clone Corp, Wuhan, China, SEC300Mu).

    Techniques: Expressing, Immunohistochemistry, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Labeling, Immunofluorescence

    Myeloid cell-specific MANF knockout promotes CCl 4 -induced HSCs activation. (A–C) The levels of α -SMA, TGF- β 1, MMP2, MMP9, TIMP1, and TIMP2 in fibrotic liver tissues were detected by using immunohistochemistry (A), Western blot (B), and qPCR (C). (D, E) Serum TGF- β 1 (D) and hyaluronan (E) levels were detected by ELISA. Data are expressed as mean ± SEM, n = 10; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. WT, wild type; HKO, MANF knockout in hepatocytes; MKO, MANF knockout in myeloid cells; CCl 4 , carbon tetrachloride.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: MANF brakes TLR4 signaling by competitively binding S100A8 with S100A9 to regulate macrophage phenotypes in hepatic fibrosis

    doi: 10.1016/j.apsb.2023.07.027

    Figure Lengend Snippet: Myeloid cell-specific MANF knockout promotes CCl 4 -induced HSCs activation. (A–C) The levels of α -SMA, TGF- β 1, MMP2, MMP9, TIMP1, and TIMP2 in fibrotic liver tissues were detected by using immunohistochemistry (A), Western blot (B), and qPCR (C). (D, E) Serum TGF- β 1 (D) and hyaluronan (E) levels were detected by ELISA. Data are expressed as mean ± SEM, n = 10; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. WT, wild type; HKO, MANF knockout in hepatocytes; MKO, MANF knockout in myeloid cells; CCl 4 , carbon tetrachloride.

    Article Snippet: The utilized ELISA Kits included Mouse TGF- β 1 ELISA Kit (R&D Systems, Minneapolis, USA, MB100B), Mouse TNF- α ELISA Kit (Abcam, ab208348), Mouse IL-1 β ELISA Kit (Abcam, ab197742), Mouse CCL2 ELISA Kit (Abcam, ab208979), Mouse S100A8 ELISA Kit (Abcam, ab263886), Mouse S100A9 ELISA Kit (Abcam, ab213887), Mouse Hyaluronan ELISA Kit (Jiyinmei Biological Technology, Wuhan, China, JYM0514Mo), and Mouse MANF ELISA Kit (Cloud-Clone Corp, Wuhan, China, SEC300Mu).

    Techniques: Knock-Out, Activation Assay, Immunohistochemistry, Western Blot, Enzyme-linked Immunosorbent Assay

    Myeloid MANF knockout promotes Ly6C high macrophages recruitment in liver. (A) Myeloid MANF deficiency increased the proportion of hepatic CD11b high F4/80 low Ly6C high mono-macrophages detected by flow cytometry. (B, C) The quantitative data in vehicle-treated (B) and CCl 4 -treated (C) mice were calculated. (D, E) The expressions of CCL2 and CX3CL1 were detected by immunohistochemistry (D) and qPCR (E). The integral optical density was calculated. (F–H) Myeloid MANF deficiency increased serum TNF- α (F), IL-1 β (G), and CCL2 (H) levels detected by ELISA. Data are expressed as mean ± SEM, n = 10; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. WT, wild type; HKO, MANF knockout in hepatocytes; MKO, MANF knockout in myeloid cells; CCl 4 , carbon tetrachloride.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: MANF brakes TLR4 signaling by competitively binding S100A8 with S100A9 to regulate macrophage phenotypes in hepatic fibrosis

    doi: 10.1016/j.apsb.2023.07.027

    Figure Lengend Snippet: Myeloid MANF knockout promotes Ly6C high macrophages recruitment in liver. (A) Myeloid MANF deficiency increased the proportion of hepatic CD11b high F4/80 low Ly6C high mono-macrophages detected by flow cytometry. (B, C) The quantitative data in vehicle-treated (B) and CCl 4 -treated (C) mice were calculated. (D, E) The expressions of CCL2 and CX3CL1 were detected by immunohistochemistry (D) and qPCR (E). The integral optical density was calculated. (F–H) Myeloid MANF deficiency increased serum TNF- α (F), IL-1 β (G), and CCL2 (H) levels detected by ELISA. Data are expressed as mean ± SEM, n = 10; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. WT, wild type; HKO, MANF knockout in hepatocytes; MKO, MANF knockout in myeloid cells; CCl 4 , carbon tetrachloride.

    Article Snippet: The utilized ELISA Kits included Mouse TGF- β 1 ELISA Kit (R&D Systems, Minneapolis, USA, MB100B), Mouse TNF- α ELISA Kit (Abcam, ab208348), Mouse IL-1 β ELISA Kit (Abcam, ab197742), Mouse CCL2 ELISA Kit (Abcam, ab208979), Mouse S100A8 ELISA Kit (Abcam, ab263886), Mouse S100A9 ELISA Kit (Abcam, ab213887), Mouse Hyaluronan ELISA Kit (Jiyinmei Biological Technology, Wuhan, China, JYM0514Mo), and Mouse MANF ELISA Kit (Cloud-Clone Corp, Wuhan, China, SEC300Mu).

    Techniques: Knock-Out, Flow Cytometry, Immunohistochemistry, Enzyme-linked Immunosorbent Assay

    MANF deficiency in myeloid cells activates S100A8/A9-TLR4 signal pathway in mice with liver fibrosis. (A) MANF deficiency in myeloid cells upregulated hepatic S100A8, S100A9, TLR4 and p-p65 levels detected by using immunohistochemistry assay. (B) The quantitative data in panel A. (C) S100A8, S100A9, TLR4, p-p65 and p65 were detected by Western blot. (D) The quantitative data in panel C. (E) S100a8 , S100a9 , and Tlr4 mRNA levels were detected by qPCR. (F) MANF deficiency in myeloid cells increased serum S100A8 and S100A9 levels. Serum S100A8 and S100A9 levels were detected by ELISA. Data are expressed as mean ± SEM, n = 10; ∗∗ P < 0.01, ∗∗∗ P < 0.001. WT, wild type; HKO, MANF knockout in hepatocytes; MKO, MANF knockout in myeloid cells; CCl 4 , carbon tetrachloride.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: MANF brakes TLR4 signaling by competitively binding S100A8 with S100A9 to regulate macrophage phenotypes in hepatic fibrosis

    doi: 10.1016/j.apsb.2023.07.027

    Figure Lengend Snippet: MANF deficiency in myeloid cells activates S100A8/A9-TLR4 signal pathway in mice with liver fibrosis. (A) MANF deficiency in myeloid cells upregulated hepatic S100A8, S100A9, TLR4 and p-p65 levels detected by using immunohistochemistry assay. (B) The quantitative data in panel A. (C) S100A8, S100A9, TLR4, p-p65 and p65 were detected by Western blot. (D) The quantitative data in panel C. (E) S100a8 , S100a9 , and Tlr4 mRNA levels were detected by qPCR. (F) MANF deficiency in myeloid cells increased serum S100A8 and S100A9 levels. Serum S100A8 and S100A9 levels were detected by ELISA. Data are expressed as mean ± SEM, n = 10; ∗∗ P < 0.01, ∗∗∗ P < 0.001. WT, wild type; HKO, MANF knockout in hepatocytes; MKO, MANF knockout in myeloid cells; CCl 4 , carbon tetrachloride.

    Article Snippet: The utilized ELISA Kits included Mouse TGF- β 1 ELISA Kit (R&D Systems, Minneapolis, USA, MB100B), Mouse TNF- α ELISA Kit (Abcam, ab208348), Mouse IL-1 β ELISA Kit (Abcam, ab197742), Mouse CCL2 ELISA Kit (Abcam, ab208979), Mouse S100A8 ELISA Kit (Abcam, ab263886), Mouse S100A9 ELISA Kit (Abcam, ab213887), Mouse Hyaluronan ELISA Kit (Jiyinmei Biological Technology, Wuhan, China, JYM0514Mo), and Mouse MANF ELISA Kit (Cloud-Clone Corp, Wuhan, China, SEC300Mu).

    Techniques: Immunohistochemistry, Western Blot, Enzyme-linked Immunosorbent Assay, Knock-Out

    MANF inhibits primary HSCs activation in vitro . (A, B) Recombinant human MANF inhibited TNF- α -induced protein (A) and mRNA (B) levels of α -SMA, TIMP1, and TIMP2 in primary HSCs detected by Western blot and qPCR, respectively. (C) Recombinant human MANF reduced TNF- α -induced α -SMA expression in HSCs. His-MANF (green) and α -SMA (red) were detected by immunofluorescent assay. DAPI (blue) was used to stain nuclei. (D) Strategy for primary HSCs treatment. The medium collected from hepatic macrophages was used to treat primary HSCs isolated from WT mice. (E) The medium collected from hepatic macrophages of MKO mice increased α -SMA level (red) in HSCs. DAPI (blue) was used to stain nuclei. (F) The quantitative data in panel E. (G, H) The medium collected from MANF-deficient macrophages increased α -SMA, TIMP1, and TIMP2 protein (G) and mRNA (H) levels of the primary HSCs detected by Western blot and qPCR, respectively. (I) MANF deficiency in hepatic macrophages increased TNF- α , IL-1 β , and CCL2 secretion in the medium detected by ELISA. (J–L) MANF deficiency in hepatic macrophages increased mRNA levels of Tnf-α (J), Il-1β (K) and Ccl2 (L) detected by qPCR. (M) Medium MANF was detected by ELISA. Data are expressed as mean ± SEM, n = 5; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. rhMANF, recombinant human MANF; WT, wild type; MKO, MANF knockout in myeloid cells; LPS, lipopolysaccharide; Mø, macrophage.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: MANF brakes TLR4 signaling by competitively binding S100A8 with S100A9 to regulate macrophage phenotypes in hepatic fibrosis

    doi: 10.1016/j.apsb.2023.07.027

    Figure Lengend Snippet: MANF inhibits primary HSCs activation in vitro . (A, B) Recombinant human MANF inhibited TNF- α -induced protein (A) and mRNA (B) levels of α -SMA, TIMP1, and TIMP2 in primary HSCs detected by Western blot and qPCR, respectively. (C) Recombinant human MANF reduced TNF- α -induced α -SMA expression in HSCs. His-MANF (green) and α -SMA (red) were detected by immunofluorescent assay. DAPI (blue) was used to stain nuclei. (D) Strategy for primary HSCs treatment. The medium collected from hepatic macrophages was used to treat primary HSCs isolated from WT mice. (E) The medium collected from hepatic macrophages of MKO mice increased α -SMA level (red) in HSCs. DAPI (blue) was used to stain nuclei. (F) The quantitative data in panel E. (G, H) The medium collected from MANF-deficient macrophages increased α -SMA, TIMP1, and TIMP2 protein (G) and mRNA (H) levels of the primary HSCs detected by Western blot and qPCR, respectively. (I) MANF deficiency in hepatic macrophages increased TNF- α , IL-1 β , and CCL2 secretion in the medium detected by ELISA. (J–L) MANF deficiency in hepatic macrophages increased mRNA levels of Tnf-α (J), Il-1β (K) and Ccl2 (L) detected by qPCR. (M) Medium MANF was detected by ELISA. Data are expressed as mean ± SEM, n = 5; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. rhMANF, recombinant human MANF; WT, wild type; MKO, MANF knockout in myeloid cells; LPS, lipopolysaccharide; Mø, macrophage.

    Article Snippet: The utilized ELISA Kits included Mouse TGF- β 1 ELISA Kit (R&D Systems, Minneapolis, USA, MB100B), Mouse TNF- α ELISA Kit (Abcam, ab208348), Mouse IL-1 β ELISA Kit (Abcam, ab197742), Mouse CCL2 ELISA Kit (Abcam, ab208979), Mouse S100A8 ELISA Kit (Abcam, ab263886), Mouse S100A9 ELISA Kit (Abcam, ab213887), Mouse Hyaluronan ELISA Kit (Jiyinmei Biological Technology, Wuhan, China, JYM0514Mo), and Mouse MANF ELISA Kit (Cloud-Clone Corp, Wuhan, China, SEC300Mu).

    Techniques: Activation Assay, In Vitro, Recombinant, Western Blot, Expressing, Staining, Isolation, Enzyme-linked Immunosorbent Assay, Knock-Out