diff --git a/django/db/models/fields/related.py b/django/db/models/fields/related.py
index 332a168a7844..56cde9cee327 100644
--- a/django/db/models/fields/related.py
+++ b/django/db/models/fields/related.py
@@ -1533,6 +1533,7 @@ def check(self, **kwargs):
             *self._check_relationship_model(**kwargs),
             *self._check_ignored_options(**kwargs),
             *self._check_table_uniqueness(**kwargs),
+            *self._check_on_delete(**kwargs),
         ]
 
     def _check_unique(self, **kwargs):
@@ -1893,6 +1894,50 @@ def _get_field_name(model):
             ]
         return []
 
+    def _check_on_delete(self, **kwargs):
+        errors = []
+        if (
+            isinstance(self.remote_field.through, str)
+            or not self.remote_field.through._meta.auto_created
+        ):
+            # Manually created through models are checked on their own.
+            return []
+        # Database and Python cascade variants cannot be mixed in a chain of
+        # model references. Auto-created through models are using Python
+        # variants.
+        m2m_through_remote_fields = (
+            m2m_model_field.remote_field
+            for m2m_model_field in self.remote_field.through._meta.get_fields()
+            if m2m_model_field.remote_field
+            and not isinstance(m2m_model_field.remote_field.model, str)
+        )
+        ref_model_fields = (
+            ref_model_field
+            for remote_field in m2m_through_remote_fields
+            for ref_model_field in remote_field.model._meta.get_fields()
+            if ref_model_field.related_model
+            and hasattr(ref_model_field.remote_field, "on_delete")
+        )
+        for ref_model_field in ref_model_fields:
+            if (
+                ref_model_field.remote_field.on_delete is not None
+                and ref_model_field.remote_field.on_delete != DO_NOTHING
+                and isinstance(ref_model_field.remote_field.on_delete, DatabaseOnDelete)
+            ):
+                errors.append(
+                    checks.Error(
+                        "Field specifies database-level on_delete variant, but "
+                        "auto-created intermediary model uses Python-level variant.",
+                        hint=(
+                            "Use either one of the Python on_delete variants or "
+                            f"create a through model for {self}."
+                        ),
+                        obj=ref_model_field,
+                        id="fields.E323",
+                    )
+                )
+        return errors
+
     def deconstruct(self):
         name, path, args, kwargs = super().deconstruct()
         # Handle the simpler arguments.
diff --git a/docs/ref/checks.txt b/docs/ref/checks.txt
index 68d1e49b1df1..14dc374b5763 100644
--- a/docs/ref/checks.txt
+++ b/docs/ref/checks.txt
@@ -322,7 +322,8 @@ Related fields
 * **fields.E322**: Field specifies ``on_delete=DB_SET_DEFAULT``, but has no
   ``db_default`` value.
 * **fields.E323**: Field specifies database/Python-level on_delete variant, but
-  referenced model uses python/database-level variant.
+  referenced/auto-created intermediary model uses python/database-level
+  variant.
 * **fields.E324**: ``<database>`` does not support
   ``<database delete option>``.
 * **fields.E330**: ``ManyToManyField``\s cannot be unique.
diff --git a/docs/releases/6.1.1.txt b/docs/releases/6.1.1.txt
index d5dbef5e790d..f53eb8c68de3 100644
--- a/docs/releases/6.1.1.txt
+++ b/docs/releases/6.1.1.txt
@@ -16,3 +16,7 @@ Bugfixes
 * Fixed a regression in Django 6.1 where :attr:`.ModelAdmin.list_display`
   entries that traverse multiple relations using ``__`` could crash or display
   incorrect values (:ticket:`37270`).
+
+* Fixed a bug in Django 6.1 where the ``fields.E323`` system check did not
+  detect mixed ``on_delete`` variants for auto-created intermediate models for
+  ``ManyToManyField``\s (:ticket:`37254`).
diff --git a/tests/invalid_models_tests/test_relative_fields.py b/tests/invalid_models_tests/test_relative_fields.py
index a6edcc4e048e..24409e9f4d95 100644
--- a/tests/invalid_models_tests/test_relative_fields.py
+++ b/tests/invalid_models_tests/test_relative_fields.py
@@ -2500,6 +2500,48 @@ class Child(models.Model):
             ],
         )
 
+    @skipUnlessDBFeature("supports_on_delete_db_cascade")
+    def test_db_python_m2m_chain(self):
+        class OtherModelParent(models.Model):
+            pass
+
+        class OtherModel(models.Model):
+            parent = models.ForeignKey(OtherModelParent, on_delete=models.DB_CASCADE)
+
+        class Parent(models.Model):
+            pass
+
+        class Child(models.Model):
+            parent = models.ForeignKey(Parent, on_delete=models.DB_CASCADE)
+            other_models = models.ManyToManyField(OtherModel)
+
+        field = Child._meta.get_field("other_models")
+        self.assertEqual(
+            field.check(from_model=Child, databases=self.databases),
+            [
+                Error(
+                    "Field specifies database-level on_delete variant, but "
+                    "auto-created intermediary model uses Python-level variant.",
+                    hint=(
+                        "Use either one of the Python on_delete variants or create a "
+                        "through model for invalid_models_tests.Child.other_models."
+                    ),
+                    obj=Child._meta.get_field("parent"),
+                    id="fields.E323",
+                ),
+                Error(
+                    "Field specifies database-level on_delete variant, but "
+                    "auto-created intermediary model uses Python-level variant.",
+                    hint=(
+                        "Use either one of the Python on_delete variants or create a "
+                        "through model for invalid_models_tests.Child.other_models."
+                    ),
+                    obj=OtherModel._meta.get_field("parent"),
+                    id="fields.E323",
+                ),
+            ],
+        )
+
     @skipUnlessDBFeature("supports_on_delete_db_cascade")
     def test_db_python_chain_auto_created(self):
         class GrandParent(models.Model):