Journal: International Journal of Molecular Sciences

Article Title: Inflammation-Driven Secretion Potential Is Upregulated in Osteoarthritic Fibroblast-Like Synoviocytes

doi: 10.3390/ijms231911817

Figure Lengend Snippet: Growth factor secretion levels by HFLS and HFLS-OA stimulated with TNFα or LPS measured using the ELISA method. Secretion of growth factors measured in media using the ELISA method. Cells were stimulated for 24 h with 10 ng/mL of TNFα ( A , C , E ) or LPS ( B , D , F ). Afterward, the media were collected, and ANGPTL1 ( A , B ), FGF5 ( C , D ), and IGF2 ( E , F ) levels were determined. Data are presented as pg or ng/mL ± SEM and analyzed with two-way ANOVA followed by Tukey’s post hoc test. *** p < 0.001 vs. control group. ## 0.01 > p > 0.001; ### p < 0.001 HFLS vs. HFLS-OA stimulated groups. HFLS, human fibroblast-like synoviocytes; HFLS-OA, osteoarthritic human fibroblast-like synoviocytes; TNFα, tumor necrosis factor alpha; LPS, lipopolysaccharide; ELISA, enzyme linked immunosorbent assay; ANGPTL1, angiopoietin like 1; FGF5, fibroblast growth factor 5; IGF2, insulin growth factor 2.

Article Snippet: The protein levels of the CXCL6 (Human GCP2 (Granulocyte Chemotactic Protein 2) ELISA Kit, ELK Biotechnology, Wuhan, China), CXCL10 (Human IP-10/CXCL10 ELISA Kit, Elabscience, Wuhan, China), CXCL16 (Human CXCL16 ELISA Kit, Elabscience, Wuhan, China), ANGPTL1 (Human ANGPTL1 ELISA Kit, ELK Biotechnology, Wuhan, China), FGF5 (Human FGF5 ELISA Kit, ELK Biotechnology, Wuhan, China) and IGF-2 (Human IGF-2 ELISA Kit, Elabscience, Wuhan, China) were measured using commercially available enzyme-linked immunosorbent assay kits according to the manufacturers’ instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 overexpression prevents SCI in mice. (A) Mice were exposed to Ctrl or SCI surgery, and FGF5 mRNA level in the spinal cord was detected at indicating times. (B) FGF5 level was detected using an ELISA kit. (C, D) Mice were intraspinally injected with 2 μL lentivirus carrying FGF5 to overexpress FGF5 in the spinal cord, and FGF5 mRNA and protein levels were detected 2 weeks post‐injection. (E) Mice were overexpressed with FGF5 using lentiviral vectors and then were exposed to SCI or sham surgery 2 weeks post‐injection. BMS score, from 0 (no ankle movement) to 9 (normal gait), was determined at indicating times. (F, G) Sensitivities to mechanical and thermal stimulation were determined 28 days after SCI. (H) Extravasation of Evans blue dye was determined 28 days after SCI. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Over Expression, Enzyme-linked Immunosorbent Assay, Injection, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 knockdown exacerbates SCI in mice. (A, B) Mice were intraspinally injected with 2 μL lentivirus carrying shFGF5 to knockdown FGF5 in the spinal cord, and FGF5 mRNA and protein levels were detected 2 weeks post‐injection. (C) Mice were injected with shFGF5, and then were exposed to SCI or sham surgery 2 weeks post‐injection. BMS score, from 0 (no ankle movement) to 9 (normal gait), was determined at indicating times. (D, E) Sensitivities to mechanical and thermal stimulation were determined 28 days after SCI. (F) Extravasation of Evans blue dye was determined 28 days after SCI. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Knockdown, Injection, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 overexpression reduces inflammation and oxidative stress in SCI mice. (A) Mice were overexpressed with FGF5 using lentiviral vectors, and then were exposed to SCI or sham surgery 2 weeks post‐injection. IL‐6 and TNF‐α levels in the spinal cord were detected. (B) MPO activity in the spinal cord. (C) Western blot images and quantification of p65 phosphorylation. (D) Relative NF‐κB transcription activity. (E, F) Relative levels of NRF2 protein and transcription activity. (G) ROS level detected by DCFH‐DA probe. (H) Quantification of hydrogen peroxide and superoxide anion in spinal cord. (I) The levels of MDA, 3‐NT and 8‐OHdG. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Over Expression, Injection, Activity Assay, Western Blot, Phospho-proteomics, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 knockdown elevates inflammation and oxidative stress in SCI mice. (A) Mice were injected with shFGF5, and then were exposed to SCI or sham surgery 2 weeks post‐injection. IL‐6 and TNF‐α levels in the spinal cord were detected. (B) MPO activity in the spinal cord. (C) Western blot images and quantification of p65 phosphorylation. (D) Relative NF‐κB transcription activity. (E, F) Relative levels of NRF2 protein and transcription activity. (G) ROS level detected by DCFH‐DA probe. (H) Quantification of hydrogen peroxide and superoxide anion in spinal cord. (I) The levels of MDA, 3‐NT and 8‐OHdG. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Knockdown, Injection, Activity Assay, Western Blot, Phospho-proteomics, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 overexpression reduces inflammation, oxidative stress and SCI through activating AMPK. (A) Mice were overexpressed with FGF5 using lentiviral vectors, and then were exposed to SCI or sham surgery 2 weeks post‐injection. Western blot images and quantification of AMPK phosphorylation in the spinal cord were detected. (B) To inhibit AMPK, mice with or without FGF5 overexpression were intraperitoneally injected with CC every 2 days 1 week pre‐SCI. IL‐6 and TNF‐α levels in the spinal cord were detected. (C) ROS level detected by DCFH‐DA probe. (D) The levels of MDA, 3‐NT and 8‐OHdG. (E) BMS score, from 0 (no ankle movement) to 9 (normal gait), was determined at indicating times. (F, G) Sensitivities to mechanical and thermal stimulation were determined 28 days after SCI. (H) Extravasation of Evans blue dye was determined 28 days after SCI. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Over Expression, Injection, Western Blot, Phospho-proteomics, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: FGF5 overexpression activates AMPK through cAMP/PKA pathway. (A, B) Mice were intraspinally injected with 2 μL lentivirus carrying FGF5 to overexpress FGF5 in the spinal cord, and cAMP level and PKA activity were detected 2 weeks post‐injection. (C) To inhibit PKA, mice with or without FGF5 overexpression were intraperitoneally injected with H89 every 2 days 1 week pre‐SCI. Western blot images and quantification of AMPK phosphorylation in the spinal cord were detected. (D) ROS level detected by DCFH‐DA probe. (E) The levels of MDA, 3‐NT and 8‐OHdG. (F) IL‐6 and TNF‐α levels in the spinal cord were detected. (G) BMS score, from 0 (no ankle movement) to 9 (normal gait), was determined at indicating times. (H, I) Sensitivities to mechanical and thermal stimulation were determined 28 days after SCI. (J) Extravasation of Evans blue dye was determined 28 days after SCI. n = 6 for each groups. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Over Expression, Injection, Activity Assay, Western Blot, Phospho-proteomics, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 5 protects against spinal cord injury through activating AMPK pathway

doi: 10.1111/jcmm.17934

Figure Lengend Snippet: Serum FGF5 level positively correlates with the sensory and motor function in SCI patients. (A) Serum levels of FGF5 in Ctrl and SCI patients. (B, C) SCI patients were divided into four groups according to serum FGF5 levels, and ASIA sensation and motor scores were measured at indicating groups. n = 56 for Ctrl group and n = 92 for SCI group. Data are expressed as the mean ± standard deviation and p < 0.05 was considered statistically significant. * p < 0.05.

Article Snippet: FGF5 in mouse spinal cord and human serum samples were detected using Mouse FGF5 ELISA Kit (#CSB‐EL008632MO; CUSABIO) or Human FGF5 ELISA Kit (#CSB‐EL008632HU; CUSABIO), respectively according to the manufacturer's instructions.

Techniques: Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Inflammation-Driven Secretion Potential Is Upregulated in Osteoarthritic Fibroblast-Like Synoviocytes

doi: 10.3390/ijms231911817

Figure Lengend Snippet: Growth factor secretion levels by HFLS and HFLS-OA stimulated with TNFα or LPS measured using the ELISA method. Secretion of growth factors measured in media using the ELISA method. Cells were stimulated for 24 h with 10 ng/mL of TNFα ( A , C , E ) or LPS ( B , D , F ). Afterward, the media were collected, and ANGPTL1 ( A , B ), FGF5 ( C , D ), and IGF2 ( E , F ) levels were determined. Data are presented as pg or ng/mL ± SEM and analyzed with two-way ANOVA followed by Tukey’s post hoc test. *** p < 0.001 vs. control group. ## 0.01 > p > 0.001; ### p < 0.001 HFLS vs. HFLS-OA stimulated groups. HFLS, human fibroblast-like synoviocytes; HFLS-OA, osteoarthritic human fibroblast-like synoviocytes; TNFα, tumor necrosis factor alpha; LPS, lipopolysaccharide; ELISA, enzyme linked immunosorbent assay; ANGPTL1, angiopoietin like 1; FGF5, fibroblast growth factor 5; IGF2, insulin growth factor 2.

Article Snippet: The protein levels of the CXCL6 (Human GCP2 (Granulocyte Chemotactic Protein 2) ELISA Kit, ELK Biotechnology, Wuhan, China), CXCL10 (Human IP-10/CXCL10 ELISA Kit, Elabscience, Wuhan, China), CXCL16 (Human CXCL16 ELISA Kit, Elabscience, Wuhan, China), ANGPTL1 (Human ANGPTL1 ELISA Kit, ELK Biotechnology, Wuhan, China), FGF5 (Human FGF5 ELISA Kit, ELK Biotechnology, Wuhan, China) and IGF-2 (Human IGF-2 ELISA Kit, Elabscience, Wuhan, China) were measured using commercially available enzyme-linked immunosorbent assay kits according to the manufacturers’ instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Inflammation-Driven Secretion Potential Is Upregulated in Osteoarthritic Fibroblast-Like Synoviocytes

doi: 10.3390/ijms231911817

Figure Lengend Snippet: Chemokine secretion levels by HFLS and HFLS-OA stimulated with TNFα or LPS measured using the ELISA method. Upregulation of chemokine level secretion measured in media using the ELISA method. Cells were stimulated for 24 h with 10 ng/mL of TNFα ( A , C , E ) or LPS ( B , D , F ). Afterward, the media were collected, and CXCL6 ( A , B ), CXCL10 ( C , D ), and CXCL16 ( E , F ) levels were determined. Data are presented as pg/mL ± SEM and analyzed with two-way ANOVA followed by Tukey’s post hoc test. ** 0.01 > p > 0.001; *** p < 0.001 vs. control group. ## 0.01 > p > 0.001; ### p < 0.001 HFLS vs. HFLS-OA stimulated groups. HFLS, human fibroblast-like synoviocytes; HFLS-OA, osteoarthritic human fibroblast-like synoviocytes; TNFα, tumor necrosis factor alpha; LPS, lipopolysaccharide; ELISA, enzyme linked immunosorbent assay; CXCL6, chemokine (C-X-C motif) ligand 6; CXCL10, chemokine (C-X-C motif) ligand 10; CXCL16, chemokine (C-X-C motif) ligand 16.

Article Snippet: The protein levels of the CXCL6 (Human GCP2 (Granulocyte Chemotactic Protein 2) ELISA Kit, ELK Biotechnology, Wuhan, China), CXCL10 (Human IP-10/CXCL10 ELISA Kit, Elabscience, Wuhan, China), CXCL16 (Human CXCL16 ELISA Kit, Elabscience, Wuhan, China), ANGPTL1 (Human ANGPTL1 ELISA Kit, ELK Biotechnology, Wuhan, China), FGF5 (Human FGF5 ELISA Kit, ELK Biotechnology, Wuhan, China) and IGF-2 (Human IGF-2 ELISA Kit, Elabscience, Wuhan, China) were measured using commercially available enzyme-linked immunosorbent assay kits according to the manufacturers’ instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Cancer cell

Article Title: Three subtypes of lung cancer fibroblasts define distinct therapeutic paradigms.

doi: 10.1016/j.ccell.2021.09.003

Figure Lengend Snippet: Figure 6. Intrinsic TGF-b signaling contributes to CAF functional heterogeneity by suppressing HGF and FGF7 expression (A) Heatmap with unsupervised clustering showing the top 1,000 differentially expressed genes across a total of 21 PDFs. (B) Volcano plots show the overexpressed genes (red) and underexpressed genes (blue) in subtype I or II PDFs compared with subtype III PDFs. (C) Venn diagram showing pathways (KEGG annotation) related with genes overexpressed in subtypes I and II PDFs. (D) RNA-seq expressions of TGF-b1 and TGF-b1 upstream suppressors DCN, FMOD, and LTBP1 (schematics on the left) in subtypes I (red), II (green), and III PDFs (blue). Mean with 95% CI. Two-tailed t test is used.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER SH-4-54 Selleckchem Cat# S7337 GDC0941 Selleckchem Cat# S1065 R406 Selleckchem Cat# S2194 SB431542 Selleckchem Cat# S1067 ABT263 Selleckchem Cat# S1001 AEW541 Selleckchem Cat# S1034 AZD0530 Selleckchem Cat# S1006 LEE011 Selleckchem Cat# S7440 MLN8237 Selleckchem Cat# S1133 Ruxolitinib Selleckchem Cat# S1378 TAE226 Selleckchem Cat# S2820 TP0903 Selleckchem Cat# S7846 Rapamycin Selleckchem Cat# S1039 Trametinib Selleckchem Cat# S2673 Vactosertib Selleckchem Cat# S7530 Cilengitide MedChemExpress Cat# HY-16141 HGF Peprotech Cat# 100-39 FGF1 Peprotech Cat# 100-17A FGF2 Peprotech Cat# 100-18C FGF5 Peprotech Cat# 100-34 FGF7 Peprotech Cat# 100-19 FGF9 Peprotech Cat# 100-23 TGF-b1 Peprotech Cat# 100-21 IL-2 STEMCELL Cat# 78036.1 DNase I STEMCELL Cat# 07900 Type-I collagen Nitta Gelatin Cat# 631-00651 Liberase Roche Cat# LIBDH-RO Blasticidin Gibico Cat# A1113903 Critical commercial assays Tumor Dissociation Kit, human Miltenyi Biotec Cat# 130-095-929 Anti-fibroblast microbeads, human Miltenyi Biotec Cat# 130-050-601 pLenti6/V5 Directional TOPO Cloning Kit Invitrogen Cat# K495510 ViraPower Lentiviral Directional TOPO Expression Kit Invitrogen Cat# K495000 Lenti-X Concentrator TaKaRa Cat# 631232 3D Cell Culture Chips AIMbiotech Cat# DAX-1, HOL-1 CellTracker Green CMFDA Dye Invitrogen Cat# C7025 CellTracker Red CMTPX Dye Invitrogen Cat# C34552 Hoechst 33342 ThermoScientific Cat# H3570 BioCoat Transwell (0.4 mm) Corning Cat# 354570 RNeasy Micro Kit Qiagen Cat# 74004 TRAPeze Telomerase Detection Kit Millipore Cat# S7700 Nunc Lab-Tek Chamber Slide System ThermoScientific Cat# 177429PK RNAscope 2.5 HD Detection Kit-Brown Advanced Cell Diagnostics Cat# 322370 RNAscope 2.5 HD Duplex Reagent Kit Advanced Cell Diagnostics Cat# 322430 Nuclear Extraction Kit Abcam Cat# ab113474 ELISA tests for HGF, FGF7 RayBiotech Cat# ELH-HGF-1, EHFGF7 ELISA tests for IGFBP6 Invitrogen Cat# EHIGFBP6 Customized quantitative proteomics array RayBiotech NA (Continued on next page) Cancer Cell 39, 1531–1547.e1–e10, November 8, 2021 e2

Techniques: Functional Assay, Expressing, RNA Sequencing, Two Tailed Test

Journal: Cancer cell

Article Title: Three subtypes of lung cancer fibroblasts define distinct therapeutic paradigms.

doi: 10.1016/j.ccell.2021.09.003

Figure Lengend Snippet: Figure 7. CAF functional classification correlates with patients’ clinical outcome (A) Normalized FGF7 and HGF secretion in 12 tumor secretome samples derived from EGFR+ NSCLC biopsies before covalent EGFR TKI (osimertinib or equivalent) treatment. Results are compared on the basis of patients’ clinical response, progressive disease (PD)/stable disease (SD) versus partial response (PR). Mean with 95% CI, one-tailed Mann-Whitney U test. (B) Functional subtypes of PDFs established from 13 NSCLC patients before receiving covalent EGFR TKI treatment (osimertinib or equivalent) are plotted against patients’ response to their treatment. (C) RNA-seq data of pre-osimertinib biopsies from 11 EGFR+ NSCLC patients (from Roper et al., 2020). The HGF and FGF7 RNA levels are based on patients’ progression-free survival (PFS) on the treatment. Mean and 95% CI are shown, one-tailed t test. (D) RNA expression of HGF and FGF7 are stained by RNAscope in pre- and post-treatment biopsy samples from two patients. (E) Functional heterogeneity in a collection of PDFs established from longitudinal biopsies from the same patients. PDFs are colored by functional subtypes. Bottom: the proportion of PDF subtypes according to early (a) and later biopsies (b, c). In (B) and (E), two-tailed Fisher’s exact test is used. See also Figure S7 and Table S5.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER SH-4-54 Selleckchem Cat# S7337 GDC0941 Selleckchem Cat# S1065 R406 Selleckchem Cat# S2194 SB431542 Selleckchem Cat# S1067 ABT263 Selleckchem Cat# S1001 AEW541 Selleckchem Cat# S1034 AZD0530 Selleckchem Cat# S1006 LEE011 Selleckchem Cat# S7440 MLN8237 Selleckchem Cat# S1133 Ruxolitinib Selleckchem Cat# S1378 TAE226 Selleckchem Cat# S2820 TP0903 Selleckchem Cat# S7846 Rapamycin Selleckchem Cat# S1039 Trametinib Selleckchem Cat# S2673 Vactosertib Selleckchem Cat# S7530 Cilengitide MedChemExpress Cat# HY-16141 HGF Peprotech Cat# 100-39 FGF1 Peprotech Cat# 100-17A FGF2 Peprotech Cat# 100-18C FGF5 Peprotech Cat# 100-34 FGF7 Peprotech Cat# 100-19 FGF9 Peprotech Cat# 100-23 TGF-b1 Peprotech Cat# 100-21 IL-2 STEMCELL Cat# 78036.1 DNase I STEMCELL Cat# 07900 Type-I collagen Nitta Gelatin Cat# 631-00651 Liberase Roche Cat# LIBDH-RO Blasticidin Gibico Cat# A1113903 Critical commercial assays Tumor Dissociation Kit, human Miltenyi Biotec Cat# 130-095-929 Anti-fibroblast microbeads, human Miltenyi Biotec Cat# 130-050-601 pLenti6/V5 Directional TOPO Cloning Kit Invitrogen Cat# K495510 ViraPower Lentiviral Directional TOPO Expression Kit Invitrogen Cat# K495000 Lenti-X Concentrator TaKaRa Cat# 631232 3D Cell Culture Chips AIMbiotech Cat# DAX-1, HOL-1 CellTracker Green CMFDA Dye Invitrogen Cat# C7025 CellTracker Red CMTPX Dye Invitrogen Cat# C34552 Hoechst 33342 ThermoScientific Cat# H3570 BioCoat Transwell (0.4 mm) Corning Cat# 354570 RNeasy Micro Kit Qiagen Cat# 74004 TRAPeze Telomerase Detection Kit Millipore Cat# S7700 Nunc Lab-Tek Chamber Slide System ThermoScientific Cat# 177429PK RNAscope 2.5 HD Detection Kit-Brown Advanced Cell Diagnostics Cat# 322370 RNAscope 2.5 HD Duplex Reagent Kit Advanced Cell Diagnostics Cat# 322430 Nuclear Extraction Kit Abcam Cat# ab113474 ELISA tests for HGF, FGF7 RayBiotech Cat# ELH-HGF-1, EHFGF7 ELISA tests for IGFBP6 Invitrogen Cat# EHIGFBP6 Customized quantitative proteomics array RayBiotech NA (Continued on next page) Cancer Cell 39, 1531–1547.e1–e10, November 8, 2021 e2

Techniques: Functional Assay, Derivative Assay, One-tailed Test, MANN-WHITNEY, RNA Sequencing, RNA Expression, Staining, RNAscope, Two Tailed Test